Top

BMC Musculoskeletal Disorders

Published in:

01-12-2020 | Research article

Adding exogenous biglycan or decorin improves tendon formation for equine peritenon and tendon proper cells in vitro

Authors: Monica Y. Pechanec, Tannah N. Boyd, Keith Baar, Michael J. Mienaltowski

Published in: BMC Musculoskeletal Disorders | Issue 1/2020

Abstract

Background

Tendon injuries amount to one of the leading causes of career-ending injuries in horses due to the inability for tendon to completely repair and the high reinjury potential. As a result, novel therapeutics are necessary to improve repair with the goal of decreasing leg lameness and potential reinjury. Small leucine-rich repeat proteoglycans (SLRPs), a class of regulatory molecules responsible for collagen organization and maturation, may be one such therapeutic to improve tendon repair. Before SLRP supplementation can occur in vivo, proper evaluation of the effect of these molecules in vitro needs to be assessed. The objective of this study was to evaluate the effectiveness of purified bovine biglycan or decorin on tendon proper and peritenon cell populations in three-dimensional tendon constructs.

Methods

Equine tendon proper or peritenon cell seeded fibrin three-dimensional constructs were supplemented with biglycan or decorin at two concentrations (5 nM or 25 nM). The functionality and ultrastructural morphology of the constructs were assessed using biomechanics, collagen content analysis, transmission electron microscopy (TEM), and gene expression by real time – quantitative polymerase chain reaction (RT-qPCR).

Results

SLRP supplementation affected both tendon proper and peritenon cells-seeded constructs. With additional SLRPs, material and tensile properties of constructs strengthened, though ultrastructural analyses indicated production of similar-sized or smaller fibrils. Overall expression of tendon markers was bolstered more in peritenon cells supplemented with either SLRP, while supplementation of SLRPs to TP cell-derived constructs demonstrated fewer changes in tendon and extracellular matrix markers. Moreover, relative to non-supplemented tendon proper cell-seeded constructs, SLRP supplementation of the peritenon cells showed increases in mechanical strength, material properties, and collagen content.

Conclusions

The SLRP-supplemented peritenon cells produced constructs with greater mechanical and material properties than tendon proper seeded constructs, as well as increased expression of matrix assembly molecules. These findings provide evidence that SLRPs should be further investigated for their potential to improve tendon formation in engineered grafts or post-injury.

Available only for authorised users

Tipton TE, Ray CS, Hand DR. Superficial digital flexor tendonitis in cutting horses: 19 cases (2007-2011). J Am Vet Med A. 2013;243:1162–5.CrossRef

Thorpe CT, Glegg PD, Birch HL. A review of tendon injury: why is the equine superficial digital flexor tendon most at risk? Equine Vet J. 2010;42(2):174–80.PubMedCrossRef

Jacobson E, Dart AJ, Mondori T, Horadogoda N, Jeffcott LB, Little CB, Smith MM. Focal experimental injury leads to widespread gene expression and histologic changes in equine flexor tendons. PLoS One. 2015;10(4):e0122220.PubMedCrossRef

Halper J. Connective tissue disorders in domestic animals. Adv Exp Med Biol. 2014;802:231–40.PubMedCrossRef

Mienaltowski MJ, Birk DE. Structure, physiology, and biochemistry of collagens. Adv Exp Med Biol. 2014;802:5–29.PubMedCrossRef

Dourte LM, Pathmanathan L, Mienaltowski MJ, Jawad AF, Birk DE, Soslowsky LJ. Mechanical, compositional, and structural properties of the mouse patellar tendon with changes in biglycan gene expression. J Orthop Res. 2013;31(9):1430–7.PubMedPubMedCentralCrossRef

Dunkman AA, Buckley MR, Mienaltowski MJ, Adams SM, Thomas SJ, Kumar A, Beason DP, Iozzo RV, Birk DE, Soslowsky LJ. The injury response of aged tendons in the absence of biglycan and decorin. Matrix Biol. 2014;35:232–8.PubMedCrossRef

Dunkman AA, Buckley MR, Mienaltowski MJ, Adams SM, Thomas SJ, Satchell L, Kumar A, Pathmanathan L, Beason DP, Iozzo RV, Birk DE, Soslowsky LF. Decorin expression is important for age-related changes in tendon structure and mechanical properties. Matrix Biol. 2013;32:3–13.PubMedCrossRef

Dunkman AA, Buckley MR, Mienaltowski MJ, Adams SM, Thomas SJ, Satchell L, Kumar A, Pathmanathan L, Beason DP, Iozzo RV, Birk DE, Soslowsky LF. The tendon injury response is influenced by decorin and biglycan. Annu Rev Biomed Eng. 2014;42:619–30.CrossRef

10.

Mienaltowski MJ, Dunkman AA, Buckley MR, Beason DP, Adams SM, Birk DE, Soslowsky LJ. Injury response of geriatric mouse patellar tendons. J Orthop Res. 2016;34:1256–63.PubMedPubMedCentralCrossRef

11.

Lechner BE, Lim JH, Mercado ML, Fallon JR. Developmental regulation of biglycan expression in muscle and tendon. Muscle Nerve. 2006;34(3):347–55.PubMedCrossRef

12.

Robinson KA, Sun M, Barnum CE, Weiss SN, Huegel J, Shetye SS, Lin L, Saez D, Adams SM, Iozzo RV, Soslowsky LJ, Birk DE. Decorin and biglycan are necessary for maintaining collagen fibril structure, fiber realignment, and mechanical properties of mature tendons. Matrix Biol. 2017;64:81–93.PubMedPubMedCentralCrossRef

13.

Bi Y, Ehirchiou D, Kilts TM, Inkson CA, Embree MC, Sonoyama W, Li L, Leet AI, Seo BM, Zhang L, Shi S, Young MF. Identification of tendon stem/progenitor cells and the role of the extracellular matrix in their niche. Nat Med. 2007;13:1219–27.PubMedCrossRef

14.

Frey H, Schroeder N, Manon-Jensen T, Iozzo RV, Schaefer L. Biological interplay between proteoglycans and their innate immune receptors in inflammation. FEBS J. 2013;280(10):2165–79.PubMedPubMedCentralCrossRef

15.

Ingraham JM, Hauck RM, Ehrlich HP. Is the tendon embryogenesis process resurrected during tendon healing? Plast Reconstr Surg. 2003;112(3):844–54.PubMedCrossRef

16.

Dyment NA, Liu CF, Kazemi N, Aschbacher-Smith L. The Paratenon contributes to Scleraxis-expressing cells during patellar tendon healing. PLoS One. 2013;8(3):e59944.PubMedPubMedCentralCrossRef

17.

Jelinsky SA, Archambault J, Li L, Seeherman H. Tendon-selective genes identified from rat and human musculoskeletal tissues. J Orthop Res. 2010;28(3):289–97.PubMedCrossRef

18.

Mienaltowski MJ, Adams SM, Birk DE. Regional differences in stem cell/progenitor cell populations from the mouse Achilles tendon. Tissue Eng Part A. 2013;19(1–2):199–210.PubMedCrossRef

19.

Mienaltowski MJ, Cánovas A, Fates VA, Hampton AR, Pechanec MY, Islas-Trejo A, Medrano JF. Transcriptome profiles of isolated murine Achilles tendon proper- and peritenon-derived progenitor cells. J Orthop Res. 2019;37(6):1409–18.PubMedCrossRef

20.

Mienaltowski MJ, Adams SM, Birk DE. Tendon proper- and peritenon-derived progenitor cells have unique tenogenic properties. Stem Cell Res Ther. 2014;5(4):86.PubMedPubMedCentralCrossRef

21.

Pechanec MY, Lee-Barthel A, Baar K, Mienaltowski MJ. Evaluation and optimization of a three-dimensional construct model for equine superficial digital flexor tendon. J Equine Vet Sci. 2018;71:90–7.CrossRef

22.

Paxton JZ, Grover LM, Baar K. Engineering an In vitro model of a functional ligament from bone to bone. Tissue Eng. 2010;16(11):3515–25.CrossRef

23.

Kapacee Z, Yeung CY, Lu Y, Crabtree D, Holmes DF, Kadler KE. Synthesis of embryonic tendon-like tissue by human marrow stromal/mesenchymal stem cells requires a three-dimensional environment and transforming growth factor β3. Matrix Biol. 2010;29(8):668–77.PubMedPubMedCentralCrossRef

24.

Amenta AR, Yilmaz A, Bogdanovich S, McKechnie BA, Abedi M, Khurana TS, Fallon JR. Biglycan recruits utrophin to the sarcolemma and counters dystrophic pathology in mdx mice. PNAS. 2011;108:762–7.PubMedCrossRef

25.

Csont T, Gorbe A, Bereczki E, Szunyog A, Aypar E, Toth ME, Varga ZV, Csonka C, Fulop F, Santha M, Ferdinandy P. Biglycan protects cardiomyocytes against hypoxia/reoxygenation injury: role of nitric oxide. J Mol Cell Cardiol. 2010;48(4):649–52.PubMedCrossRef

26.

Reese SP, Underwood CJ, Weiss JA. Effects of decorin proteoglycan on fibrillogenesis, ultrastructure, and mechanics of type I collagen gels. Matrix Biol. 2013;32(7–8):414–23.PubMedCrossRef

27.

Lee-Barthel A, Baar K, West DWD. Treatment of ligament constructs with exercise-conditioned serum: a translational tissue engineering model. JOVE-J Vis Exp. 2017;124:1–13.

28.

Kapacee Z, Richardson SH, Lu Y, Starborg T, Holms DF, Baar K, Kadler KE. Tension is required for Fibripositor formation. Matrix Biol. 2008;27(4):371–5.PubMedCrossRef

29.

Calve S, Dennis RG, Kosnik PE II, Baar K, Grosh K, Arruda EM. Engineering of functional tendon. Tissue Eng. 2004;10(5/6):755–61.PubMedCrossRef

30.

Larkin LM, Calve S, Kostrominova TY, Arruda EM. Structure and functional evaluation of tendon – skeletal muscle constructs engineered in Vitro. Tissue Eng. 2006;12(11):3149–58.PubMedPubMedCentralCrossRef

31.

Woessner JF Jr. The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid. Arch Biochem Biophys. 1961;93:440–7.PubMedCrossRef

32.

Edwards CA, O’Brien WD Jr. Modified assay for determination of hydroxyproline in a tissue hydrolyzate. Clin Chim Acta. 1980;104(2):161–7.PubMedCrossRef

33.

Mushran SP, Agrawal MC, Prasad B. Mechanism of oxidation by chloramine-T. Part I. Oxidation of α-hydroxy-acids. J Chem Soc. B 1971;Part I:1712–4.

34.

Creemers LB, Jansen DC, van Veen-Reurings A, Van den Bos T, Everts V. Microassay for the assessment of low levels of hydroxyproline. Biotechniques. 1997;22:656–8.PubMedCrossRef

35.

Birk DE, Trelstad RL. Extracellular compartments in matrix morphogenesis: collagen fibril, bundle, and lamellar formation by corneal fibroblasts. J Cell Bio. 1984;99:2024–33.CrossRef

36.

Birk DE, Trelstad RL. Extracellular compartments in tendon morphogenesis: collagen fibril, bundle, and macroaggregate formation. J Cell Bio. 1986;103:231–40.CrossRef

37.

Mienaltowski MJ, Huang L, Frisbie DD, McIlwraith SW, Stromberg AJ, Bathke AC, MacLeod JN. Transcriptional profiling differences for articular cartilage and repair tissue in equine joint surface lesions. BMC Med Genet. 2009;2:60.

38.

Mienaltowski MJ, Huang L, Stromberg AJ, MacLeod JN. Differential gene expression associated with postnatal equine articular cartilage maturation. BMC Musculoskelet Disord. 2008;9:149.PubMedPubMedCentralCrossRef

39.

Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 2002;3:RESEARCH0034.PubMedPubMedCentralCrossRef

40.

Scott EY, Mansour T, Bellone RR, Brown C, Mienaltowski MJ, Penedo MC, Ross PJ, Valberg SJ, Murray JD, Finno CJ. Identification of long non-coding RNA in the horse transcriptome. BMC Genomics. 2017;18(1):511.PubMedPubMedCentralCrossRef

41.

Mansour TA, Scott EY, Finno CJ, Bellone RR, Mienaltowski MJ, Pendo MC, Ross PJ, Valberg SJ, Murray JD, Brown CT. Tissue resolved, gene structure refined equine transcriptome. BMC Genomics. 2017;18(1):103.PubMedPubMedCentralCrossRef

42.

Ramakers C, Ruijter JM, Deprez RH, Moorman AF. Assumption-free analysis of quantitative real-time polymerase chain reaction (PCR) data. Neurosci Lett. 2003;339:62–6.PubMedCrossRef

43.

Schefe JH, Lehmann KE, Buschmann IR, Unger T, Funke-Kaiser H. Quantitative real-time RT-PCR data analysis: current concepts and the novel “gene expression's CT difference” formula. J Mol Med (Berl). 2006;84:901–10.CrossRef

44.

Wilcoxon F. Individual comparisons by ranking methods. Biom Bull. 1945;1(6):80–3.CrossRef

45.

Bland JM, Altman DG. One and two sided test of significance. BMJ. 1994;309(6949):248.PubMedPubMedCentralCrossRef

46.

Murphy R. On the use of one-sided statistical tests in biomedical research. Clin Exp Pharmacol Physiol. 2018;45(1):109–14.PubMedCrossRef

47.

Danielson KG, Baribault H, Holmes DF, Graham H, Kadler KE, Iozzo RV. Targeted disruption of decorin leads to abnormal collagen fibril morphology and skin fragility. J Cell Bio. 1997;136(3):729–43.CrossRef

48.

Wadhwa S, Embree MC, Kilts T, Young MF, Ameye LG. Accelerated osteoarthritis in the temporomandibular joint of biglycan/fibromodulin double-deficient mice. Osteoarthr Cartilage. 2005;13(9):817–27.CrossRef

49.

Parisuthiman D, Mochida Y, Duarte WR, Yamauchi M. Biglycan modulates osteoblast differentiation and matrix mineralization. J Bone Miner Res. 2009;20(10):1878–86.CrossRef

50.

Han B, Li Q, Wang C, Patel P, Adams SM, Doyran B, Nia HT, Oftadeh R, Zhou S, Li CY, Liu XS, XL LU, Enomoto-Iwamoto M, Qin L, Mauck RL, Iozzo RV, Birk DE, Han L. Decorin regulates the aggrecan network integrity and biomechanical functions of cartilage extracellular matrix. ACS Nano. 2019;13(10):11320–33.PubMedCrossRefPubMedCentral

51.

Delalande A, Gosselin MP, Suwalski A, Guilmain W, Leduc C, Berchel M, Jaffres PA, Baril P, Midoux P, Pichon C. Enhanced Achilles tendon haling by fibromodulin gene transfer. Nanomedicine. 2015;11(7):1735–44.PubMedCrossRef

52.

Ye Y, Hu G, Guo F, Zhang W, Wang J, Chen A. Glycosaminoglycan chains of biglycan promote bone morphogenetic protein-4-induced osteoblast differentiation. Int J Mol Med. 2012;30(5):1075–80.PubMedCrossRef

53.

Berendsen AD, Fisher LW, Kilts TM, Owens RT, Robey PG, Gutkind JS, Young MF. Modulation of canonical Wnt signaling by the extracellular matrix component biglycan. Proc Natl Acad Sci U S A. 2011;108(41):17022–7.PubMedPubMedCentralCrossRef

54.

Schonherr E, Levkau B, Schaefer L, Kresse H, Walsh K. Decorin affects endothelial cells by Akt-dependent and -independent pathways. Ann N Y Acad Sci. 2002;973:149–52.PubMedCrossRef

55.

Mohan RR, Tripathi R, Sharma A, Sinha PR, Giuliano EA, Hesemann NP, Chaurasia SS. Decorin antagonizes corneal fibroblast migration via caveolae-mediated endocytosis of epidermal growth factor receptor. Exp Eye Res. 2019;180:200–7.PubMedCrossRef

56.

Geng Y, McQuillan D, Roughley PJ. SLRP interaction can protect collagen fibrils from cleavage by collagenases. Matrix Biol. 2006;25(8):484–91.PubMedCrossRef

57.

Meij JT, Carlson EC, Wang L, Liu CY, Jester JV, Birk DE, Kao WW. Targeted expression of a lumican transgene rescues corneal deficiencies in lumican-null mice. Mol Vis. 2007;13:2012–8.PubMed

58.

Jiang W, Ting K, Lee S, Zara JN, Song R, Li C, Chen E, Zhang X, Zhao Z, Soo C, Zheng Z. Fibromodulin reduces scar size and increases scar tensile strength in normal and excessive-mechanical-loading porcine cutaneous wounds. J Cell Mol Med. 2017;22(4):2510–3.CrossRef

59.

Gaspar R, Pipicz M, Hawchar F, Kovacs D, Djirackor L, Gorbe A, Varga ZV, Kiricsi M, Ptrovski G, Gacser A, Csonka C, Csont T. The cryoprotective effect of biglycan core protein involves toll-like receptor 4 signaling in cardiomyocytes. J Mol Cell Cardiol. 2016;99:138–50.PubMedCrossRef

60.

Barnette DN, Hulin A, Ahmed AS, Colige AC, Azhar M, Lincoln J. TGFβ-Smad and MAPK signaling mediate scleraxis and proteoglycan expression in heart valves. J Mol Cell Cardiol. 2013;65:137–46.PubMedCrossRef

61.

Farhat YM, Al-Maliki AA, Chen T, Juneja SC, Schwarz EM, O’Kefe RJ, Awad HA. Gene expression analysis of the pleitrophic effects of TGF-β1 in an in vitro model of flexor tendon healing. PLoS One. 2012;7(12):e51411.PubMedPubMedCentralCrossRef

62.

Berendsen AD, Pinnow EL, Maeda A, et al. Biglycan modulates angiogenesis and bone formation during fracture healing. Matrix Biol. 2014;35:223–31.PubMedCrossRef

63.

Neill T, Painter H, Buraschi S, et al. Decorin antagonizes the angiogenic network. Concurrent inhibition of met, hypoxia inducible factor-1α and vascular endothelial growth factor a and induction of thrombospondin-1 and TIMP3. J Biol Chem. 2012;287:5492–506.PubMedCrossRef

64.

Korntner S, Lehner C, Gehwolf R, Wagner A, Grutz M, Kunkel N, Temfer H, Traweger A. Limiting angiogenesis to modulate scar formation. Adv Drug Deliv Rev. 2019;146:170–89.PubMedCrossRef

65.

DiPietro LA. Angiogenesis and scar formation in healing wounds. Curr Opin Rheumatol. 2013;25(1):87–91.PubMedCrossRef

66.

Cadby JA, Buehler E, Godbout C, van Weeren RP, Snedeker JG. Differences between the cell populations from the peritenon and the tendon core with regard to their potential implications in tendon repair. PLoS One. 2014;9(3):e92474.PubMedPubMedCentralCrossRef

67.

Costa-Almeida R, Gonçalves AI, Gershovich P, Rodrigues MT, Reis RL, Gomes ME. Tendon stem cell niche. In: Turksen K, editor. Tissue engineering and stem cell niche. Cham: Springer; 2015. Stem Cell Niche.

68.

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;9(4):e96113.PubMedPubMedCentralCrossRef

Title: Adding exogenous biglycan or decorin improves tendon formation for equine peritenon and tendon proper cells in vitro
Authors: Monica Y. Pechanec
Tannah N. Boyd
Keith Baar
Michael J. Mienaltowski
Publication date: 01-12-2020
Publisher: BioMed Central
Published in: BMC Musculoskeletal Disorders / Issue 1/2020
Electronic ISSN: 1471-2474
DOI: https://doi.org/10.1186/s12891-020-03650-2

At a glance: The STEP trials

Springer Medicine

Adding exogenous biglycan or decorin improves tendon formation for equine peritenon and tendon proper cells in vitro

Abstract

Background

Methods

Results

Conclusions

At a glance: The STEP trials

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2020

Oxygen embolism after hydrogen peroxide irrigation during hip arthroscopy: a case report

Correction to: On-site multi-component intervention to improve productivity and reduce the economic and personal burden of neck pain in Swiss office-workers (NEXpro): protocol for a cluster-randomized controlled trial

No differences in cost-effectiveness and short-term functional outcomes between cemented and uncemented total knee arthroplasty

Development and validation of the General Rehabilitation Adherence Scale (GRAS) in patients attending physical therapy clinics for musculoskeletal disorders

Association of complete uncinate process removal on 2-year assessment of radiologic outcomes: subsidence and sagittal balance in patients receiving one-level anterior cervical discectomy and fusion

Clinical indicators to identify neuropathic pain in low back related leg pain: a modified Delphi study