Objective: The aim of this study was to investigate the effect of mucopolysaccharide, vitamin C, and collagen supplementation on the healing of Achilles tendon in rats. Methods: Sixteen rats were separated into 2 groups. Both Achilles tendons of all rats were transected 5 mm above the insertion and repaired using a Kessler suture. After the surgical repair, the study group received the daily recommended amount of the supplement by gastric gavage, while the control group received a placebo. At the end of the third week, the animals were sacrificed. The biomechanical properties of the groups were compared with ultimate tensile strength and stiffness tests. The biological properties of the 2 groups were assessed with a histomorphometric comparison to determine the amount of collagen type I (COL1), proliferating cell nuclear antigen (PCNA), and transforming growth factor b1 (TGF-b1) expression in 3 different tissue subgroups (collagen matrix, tenocytes, and endotenon fibroblasts). Results: Analysis of histomorphometric results revealed that the rats receiving dietary supplements demonstrated a significant increase in PCNA (mean value of 86 in the control group and 168.85 in the trial group; p < 0.05) and TGF-b1 (mean value of 87.57 in the control group and 161.85 in the trial group; p < 0.05) in the endotenon fibroblasts of the repair site. However, there was no difference between the groups in PCNA or TGF-b1 when the collagen matrix and the tenocytes of the repair site were examined. Furthermore, no significant difference could be found between groups in COL1 in any of the 3 tissue subgroups (collagen matrix, tenocytes, and endotenon fibroblasts). The statistical analysis also indicated that the rats receiving supplements did not demonstrate a significant increase in the ultimate tendon tensile strength or stiffness. Conclusion: The results of this study revealed no advantage to the oral administration of the trial supplement in collagen synthesis or biomechanical properties in rats after 3 weeks using the presented study design. However, the increased expression of PCNA and TGFb1 seen in the endotenon fibroblasts of the repair site might play a role in the continuum of tendon healing.
___
Langberg H, Skovgaard D, Asp S, Kjaer M. Time pattern of exercise-induced changes in type I collagen turnover after prolonged endurance exercise in humans. Calcif Tissue Int. 2000 Jul;67(1):41e44. https://doi.org/10.1007/ s00223001094.
Dunning J, Butts R, Mourad F, Young I, Flannagan S, Perreault T. Dry needling: a literature review with implications for clinical practice guidelines. Phys Ther Rev. 2014 Aug;19(4):252e265. https://doi.org/10.1179/ 108331913X13844245102034.
Rabago D, Lee KS, Ryan M, et al. Hypertonic dextrose and morrhuate sodium injections (prolotherapy) for lateral epicondylosis (Tennis elbow): results of a single-blind, pilot-level randomized controlled trial. Am J Phys Med Rehabil. 2013 Jul;92(7):587e596. https://doi.org/10.1097/PHM.0b013e31827d695f.
Sanderson LM, Bryant A. Effectiveness and safety of prolotherapy injections for management of lower limb tendinopathy and fasciopathy: a systematic review. J Foot Ankle Res. 2015 Oct;8:57. https://doi.org/10.1186/s13047-015-0114-5. eCollection 2015.
Best TM, Zgierska AE, Zeisig E, Ryan M, Crane D. A systematic review of four injection therapies for lateral epicondylosis: prolotherapy, polidocanol, whole blood and platelet rich plasma. Br J Sports Med. 2009 Jul;43(7):471e481. https://doi.org/10.1136/bjsm.2008.052761.
Jeong DU, Lee CR, Lee JH, et al. Clinical applications of platelet-rich plasma in patellar tendinopathy. Biomed Res Int. 2014 Jul;2014:249498. https://doi.org/ 10.1155/2014/249498.
Merolla G, Dellabiancia F, Ingardia A, Paladini P, Porcellini G. Co-analgesic therapy for arthroscopic supraspinatus tendon repair pain using a dietary supplement containing Boswellia serrata and curcuma longa: a prospective randomized placebo-controlled study. Musculoskelet Surg. 2015 Sep;99(suppl 1):S43eS52. https://doi.org/10.1007/s12306-015-0364-1 [Epub 2015 May 10].
Notarnicola A, Pesce V, Vicenti G, Tafuri S, Forcignano M, Moretti B. SWAAT study: extracorporeal shock wave therapy and arginine supplementation and other nutraceuticals for insertional Achilles tendinopathy. Adv Ther. 2012 Sep;29(9):799e814. https://doi.org/10.1007/s12325-012-0046-4 [Epub 2012 Aug 24].
Gumina S, Passaretti D, Gurzi MD, Candela V. Arginine L-alpha-ketoglutarate, methylsulfonylmethane, hydrolyzed type I collagen and bromelain in rotator cuff tear repair: a prospective randomized study. Curr Med Res Opin. 2012 Nov;28(11):1767e1774. https://doi.org/10.1185/03007995.2012.737772 [Epub 2012 Oct 19].
Sundqvist H, Forsskahl B, Kvist M. A promising novel therapy for Achilles peritendinitis: double-blind comparison of glycosaminoglycan polysulfate and high-dose indomethacin. Int J Sports Med. 1987 Aug;8(4):298e303.
Shakibaei M, Buhrmann C, Mobasheri A. Anti-inflammatory and anti-catabolic effects of TENDOACTIVE(R) on human tenocytes in vitro. Histol Histopathol. 2011 Sep;26(9):1173e1185. https://doi.org/10.14670/HH-26.1173 .
Usha PR, Naidu MU. Randomised, double-blind, parallel, placebo-controlled study of oral glucosamine, methylsulfonylmethane and their combination in osteoarthritis. Clin Drug Invest. 2004 Jun;24(6):353e363. https://doi.org/ 10.2165/00044011-200424060-00005.
Buhrmann C, Mobasheri A, Busch F, et al. Curcumin modulates nuclear factor kappaB (NF-kappaB)-mediated inflammation in human tenocytes in vitro: role of the phosphatidylinositol 3-kinase/Akt pathway. J Biol Chem. 2011 Aug;286(32):28556e28566. https://doi.org/10.1074/jbc.M111.256180 [Epub 2011 Jun 13].
Kubaski F, Suzuki Y, Orii K, et al. Glycosaminoglycan levels in dried blood spots of patients with mucopolysaccharidoses and mucolipidoses. Mol Genet Metab. 2017 Mar;120(3):247e254. https://doi.org/10.1016/j.ymgme.2016.12.010.
Shimada T, Tomatsu S, Yasuda E, et al. Chondroitin 6-sulfate as a novel biomarker for mucopolysaccharidosis IVA and VII. JIMD Rep. 2014 May;16: 15e24. https://doi.org/10.1007/8904_2014_311. Published online 2014 May 22.
Tas¸ kesen A, Ataoglu B, Ozer M, Demirkale I, Turanli S. Glucosamine-chondroitin € sulphate accelerates tendon-to-bone healing in rabbits. Eklem Hastalik Cerrahisi. 2015;26(2):77e83. https://doi.org/10.5606/ehc.2015.17.
Oryan A, Moshiri A, Meimandiparizi AH. Effects of sodium-hyaluronate and glucosamine-chondroitin sulfate on remodeling stage of tenotomized superficial digital flexor tendon in rabbits: a clinical, histopathological, ultrastructural, and biomechanical study. Connect Tissue Res. 2010 Nov;52(4):329e339. https://doi.org/10.3109/03008207.2010.531332.
Lippiello L. Collagen synthesis in tenocytes, ligament cells and chondrocytes exposed to a combination of glucosamine HCl and chondroitin sulfate. Evid Based Complement Alternat Med. 2007 Jun;4(2):219e224. https://doi.org/ 10.1093/ecam/nel081.
Minaguchi J, Koyama Y, Meguri N, et al. Effects of ingestion of collagen peptide on collagen fibrils and glycosaminoglycans in Achilles tendon. J Nutr Sci Vitaminol (Tokyo). 2005 Jun;51(3):169e174. https://doi.org/10.3177/jnsv.51.169.
Shaw G, Lee-Barthel A, Ross ML, Wang B, Baar K. Vitamin C-enriched gelatin supplementation before intermittent activity augments collagen synthesis. Am J Clin Nutr. 2017 Jan;105(1):136e143. https://doi.org/10.3945/ajcn.116. 138594.
Sato Y, Mera H, Takahashi D, et al. Synergistic effect of ascorbic acid and collagen addition on the increase in type 2 collagen accumulation in cartilagelike MSC sheet. Cytotechnology. 2017 Jun;69(3):405e416. https://doi.org/ 10.1007/s10616-015-9924-3.
Tsai WC, Hsu CC, Chung CY, Lin MS, Li SL, Pang JH. The pentapeptide KTTKS promoting the expressions of type I collagen and transforming growth factorbeta of tendon cells. J Orthop Res. 2007 Dec;25(12):1629e1634. https://doi.org/ 10.1002/jor.20455.
Chen MH, Huang YC, Sun JS, Chao YH, Chen MH. Second messengers mediating the proliferation and collagen synthesis of tenocytes induced by low-level laser irradiation. Lasers Med Sci. 2015 Jan;30(1):263e272. https://doi.org/10.1007/ s10103-014-1658-5.
Probst A, Palmes D, Freise H, Langer M, Joist A, Spiegel HU. A new clamping technique for biomechanical testing of tendons in small animals. J Invest Surg. 2000 Nov-Dec;13(6):313e318.
Kelman Z. PCNA: structure, functions and interactions. Oncogene. 1997 Feb 13;14(6):629e640.
Prosperi E. Multiple roles of the proliferating cell nuclear antigen: DNA replication, repair and cell cycle control. In: Laurent M, Silvana G, Michel P, eds. Progress in Cell Cycle Research. vol. 3. Boston: Springer; 1997:193e210. https:// doi.org/10.1007/978-1-4615-5371-7_15.
Khan U, Occleston NL, Khaw PT, McGrouther DA. Differences in proliferative rate and collagen lattice contraction between endotenon and synovial fibroblasts. J Hand Surg [Am]. 1998 Mar;23(2):266e273. https://doi.org/10.1016/ S0363-5023(98)80125-1.
Khan U, Kakar S, Akali A, Bentley G, McGrouther DA. Modulation of the formation of adhesions during the healing of injured tendons. J Bone Joint Surg Br. 2000 Sep;82(7):1054e1058. https://doi.org/10.1302/0301-620X.82B7.0821054.
Tokunaga T, Shukunami C, Okamoto N, et al. FGF-2 stimulates the growth of tenogenic progenitor cells to facilitate the generation of tenomodulin-positive tenocytes in a rat rotator cuff healing model. Am J Sports Med. 2015 Oct;43(10): 2411e2422. https://doi.org/10.1177/0363546515597488 [Epub 2015 Aug 26].
Dyment NA, Hagiwara Y, Matthews BG, Li Y, Kalajzic I, Rowe DW. Lineage tracing of resident tendon progenitor cells during growth and natural healing. PLoS One. 2014 Apr 23;9(4):e96113. https://doi.org/10.1371/journal.pone. 0096113. eCollection 2014.
Tsubone T, Moran SL, Subramaniam M, Amadio PC, Spelsberg TC, An KN. Effect of TGF-beta inducible early gene deficiency on flexor tendon healing. J Orthop Res. 2006 Mar;24(3):569e575. https://doi.org/10.1002/jor.20101.
O'Kane S, Ferguson MW. Transforming growth factor beta s and wound healing. Int J Biochem Cell Biol. 1997 Jan;29(1):63e78. https://doi.org/10.1016/ S1357-2725(96)00120-3.
Beck LS, Deguzman L, Lee WP, et al. TGF-beta 1 accelerates wound healing: reversal of steroid-impaired healing in rats and rabbits. Growth Factors. 1991;5(4):295e304.
Tajima S, Pinnell SR. Ascorbic acid preferentially enhances type I and III collagen gene transcription in human skin fibroblasts. J Dermatol Sci. 1996 Mar;11(3):250e253. https://doi.org/10.1016/0923-1811(95)00640-0.
Kishimoto Y, Saito N, Kurita K, Shimokado K, Maruyama N, Ishigami A. Ascorbic acid enhances the expression of type 1 and type 4 collagen and SVCT2 in cultured human skin fibroblasts. Biochem Biophys Res Commun. 2013 Jan 11;430(2):579e584. https://doi.org/10.1016/j.bbrc.2012.11.110.
Omeroglu S, Peker T, Turkozkan N, Omeroglu H. High-dose vitamin C supplementation accelerates the Achilles tendon healing in healthy rats. Arch Orthop Trauma Surg. February 2009;129(2):281e286. https://doi.org/10.1007/s00402- 008-0603-0 [Epub 2008 Feb 29].
Danielski LG, Walczewski E, de Jesus CR, et al. Preoperative vitamin C supplementation improves colorectal anastomotic healing and biochemical parameters in malnourished rats. Int J Colorectal Dis. 2016 Nov;31(11):1759e1766. https:// doi.org/10.1007/s00384-016-2647-x.
Findik RB, Ilkaya F, Guresci S, Guzel H, Karabulut S, Karakaya J. Effect of vitamin C on collagen structure of cardinal and uterosacral ligaments during pregnancy. Eur J Obstet Gynecol Reprod Biol. 2016 Jun;201:31e35. https://doi.org/ 10.1016/j.ejogrb.2016.03.022.
Blobe GC, Schiemann WP, Lodish HF. Role of transforming growth factor beta in human disease. N Engl J Med. 2000 May;342(4):1350e1358. https://doi.org/ 10.1056/NEJM200005043421807.
Singer AJ, Clark RA. Cutaneous wound healing. N Engl J Med. 1999 Sep;341(10): 738e746. https://doi.org/10.1056/NEJM199909023411006.