Histology and immunohistochemistry study of ovine tendon grafted with cBMSCs and BMMNCs after collagenase-induced tendinitis

A. Crovace; L. Lacitignola; E. Francioso; G. Rossi

doi:10.3415/VCOT-07-05-0050

Veterinary and Comparative Orthopaedics and Traumatology, Table of Contents

Vet Comp Orthop Traumatol 2008; 21(04): 329-336
DOI: 10.3415/VCOT-07-05-0050

Original Research

Schattauer GmbH

Histology and immunohistochemistry study of ovine tendon grafted with cBMSCs and BMMNCs after collagenase-induced tendinitis

Authors

A. Crovace

¹Dipartimento dell’ Emergenza e dei Trapianti di Organi (D.E.T.O.), Sezione di Chirurgia Veterinaria, Facoltà di Medicina Veterinaria, Università degli Studi di Bari, Bari, Italy
L. Lacitignola

²Dipartimento di Scienze Cliniche Veterinarie, Sezione di Chirurgia Veterinaria, Facoltà di Medicina Veterinaria, Università degli Studi di Teramo, Teramo, Italy
E. Francioso

¹Dipartimento dell’ Emergenza e dei Trapianti di Organi (D.E.T.O.), Sezione di Chirurgia Veterinaria, Facoltà di Medicina Veterinaria, Università degli Studi di Bari, Bari, Italy
G. Rossi

³Dipartimento di Scienze Veterinarie, Università degli Studi di Camerino, Facoltà di Medicina Veterinaria, Matelica (MC), Italy

Abstract

Summary

Objectives: The aim of this study was to compare the regeneration abilities of cultured bone marrow mesenchymal cells (cBMSC) and bone marrow mononuclear cells (BMMNC) with fibrin glue, saline solution and sham control in collagenase–induced tendinitis of the Achilles tendon in sheep. Methods: Six sheep were recruited randomly to each group: cBMSC, BMMNC, fibrin, saline and sham control. Each group received the relative treatment two weeks after inducing lesions (T0). After eight weeks (T8) of treatment, the tendons were harvested and evaluated for histomorphology, Collagen type I, III, Cartilage Oligomeric Matrix Protein (COMP) and CD34 positive cells expression. Results: Histology and immunohistochemistry showed similar capabilities of cBMSC and BMMNC to restore the architecture of fibres and Extra Cellular Matrix (ECM), with a high expression of collagen type I and COMP and a very low expression of collagen type III in treated tendons. The complete architectural disruption of fibres, dramatic reduction of collagen Type I and COMP expression and increase collagen type III expression were commonly observed in tendons treated with fibrin or saline only. The presence of CD34 positive cells was appreciable in the BMMNC group while few cBMSC showed this cluster of differentiation, not expressed in tendons treated with fibrin or saline. Clinical significance: The data in this study show the efficacy of cBMSC and BMMNC in regenerating tendon tissue after collagenase–induced tendinitis.

Keywords

Collagenase tendonitis - cultured bone marrow mesenchymal stem cells - bone marrow mononuclear cells - autograft - achilles tendon - sheep

Full Text

References

References
1 Caplan AI, Bruder SP. Mesenchymal stem cells: building blocks formolecularmedicine in the 21st century.. Trends Mol Med 2001; 7: 259-264.
2 Fortier LA. Stem cells: classifications, controversies, and clinical applications.. Vet Surg 2005; 34: 415-423.
3 Awad HA, Butler DL, Boivin GP. et al. Autologous mesenchymal stem cell-mediated repair of tendon.. Tissue Eng 1999; 5: 267-277.
4 Awad HA, Butler DL, Harris MT. et al. In vitro characterization of mesenchymal stem cell- seeded collagen scaffolds for tendon repair: effects of initial seeding density on contraction kinetics.. J Biomed Mater Res 2000; 51: 233-240.
5 Awad HA, Boivin GP, Dressler MR. et al. Repair of patellar tendon injuries using a cell-collagen composite.. J Orthop Res 2003; 21: 420-431.
6 Ouyang HW, Goh JC, Thambyah A. et al. Knitted poly-lactide-co-glycolide scaffold loaded with bone marrow stromal cells in repair and regeneration of rabbit Achilles tendon.. Tissue Eng 2003; 9: 431-439.
7 Ouyang HW, Goh JC, Lee EH. Viability of allogeneic bone marrow stromal cells following local delivery into patella tendon in rabbit model.. Cell Transplant 2004; 13: 649-657.
8 Ouyang HW, Goh JC, Lee EH. Use of bone marrow stromal cells for tendon graft-to-bone healing: histological and immunohistochemical studies in a rabbit model.. Am J Sports Med 2004; 32: 321-327.
9 Pittenger M, Vanguri P, Simonetti D. et al. Adult mesenchymal stem cells: potential formuscle and tendon regeneration and use in gene therapy.. J Musculoskelet Neuronal Interact 2002; 2: 309-320.
10 Young RG, Butler DL, Weber W. et al. Use of mesenchymal stem cells in a collagen matrix for Achilles tendon repair.. J Orthop Res 1998; 16: 406-413.
11 Smith RK, Korda M, Blunn GW. et al. Isolation and implantation of autologous equine mesenchymal stem cells from bone marrow into the superficial digital flexor tendon as a potential novel treatment.. Equine Vet J 2003; 35: 99-102.
12 Smith RK. Autogenous Stem Cell Implantation.. Proceedings of ACVS Veterinary Symposium, October 7-9, 2004, Denver, CO. Vet Surg 2004; 33: 199-201 E.
13 Smith RK, Webbon PM. Harnessing the stem cell for the treatment of tendon injuries: heralding a new dawn?. Br J Sports Med 2005; 39: 582-584.
14 Smith RK. The use of bone marrow-derived stem cells in tendon injury. American College of Veterinary Surgeons Symposium October 5, 2006; 155-156.
15 Hisatome T, Yasunaga Y, Yanada S. et al. Neovascularization and bone regeneration by implantation of autologous bone marrow mononuclear cells.. Biomaterials 2005; 26: 4550-4556.
16 Lin GS, Lu JJ, Jiang XJ. et al. Autologous transplantation of bone marrow mononuclear cells improved heart function after myocardial infarction.. Acta Pharmacol Sin 2004; 25: 876-886.
17 Ryu JH, Kim IK, Cho SW. et al. Implantation of bone marrow mononuclear cells using injectable fibrin matrix enhances neovascularization in infarcted myocardium.. Biomaterials 2005; 26: 319-326.
18 Tateishi-Yuyama E, Matsubara H, Murohara T. et al. Therapeutic angiogenesis for patients with limb ischaemia by autologous transplantation of bone-marrow cells: apilot study and arandomised controlled trial.. Lancet 2002; 360: 427-435.
19 Tse HF, Kwong YL, Chan JK. et al. Angiogenesis in ischaemic myocardium by intramyocardial autologous bone marrow mononuclear cell implantation.. Lancet 2003; 361: 47-49.
20 Waksman R, Balfour R. Bone marrow and bone marrow derived mononuclear stem cells therapy for the chronically ischemic myocardium.. Cardiovasc Radiat Med 2003; 4: 164-168.
21 Crovace A, Lacitignola L, De Siena R. et al. La rigenerazione del tendine nel cavallo mediante terapia cellulare.. Studio sperimentale; Società Italiana Delle Scienze Veterinarie Lx Convegno Nazionale. 327-328 September 27 2006 .
22 Crovace A, Lacitignola L, De Siena R. et al. Bone marrowmononucleated cells (BMMNCs) are able to synthesize type I collagen similar to cultured bone marrow stromal cells (cBMSC) in an equine model of collagenase-induced tendonitis: A preliminary study; CESMAS 2006: Management of lameness causes in sport horses. 2006; July 19 169-172.
23 Crovace A, Lacitignola L, De Siena R. et al. Cultured bone marrow stromal cells, bone marrow mononucleated cells, and placebo in experimental tendinitis in the horse: a preliminary study; 4^th ISSCR Annual Meeting. 2006; June 29 87.
24 Crovace A, Francioso E, Hendry J. et al. Use of autogenous bone marrow stromal cells (BMSC) and resorbable scaffolds in dogs with bone loss. 12^th ESVOT Congress, 10^th-12^th September, Munich, 2004, published in. Vet Comp Orthop Traumatol 2004; 18: A27.
25 Crovace A, Francioso E, Hendry J. et al. Use of autologous bone marrow stromal cell (BMSC) and resorbable bone graft substitute in dogs with orthopedic lesions.. Proceedings of ACVS Veterinary Symposium, October 7-9, 2004, Denver, CO. Vet Surg 2004; 33: 435-E25.
26 Crovace A, Lacitignola L, De Siena R. et al. Trattamento delle lesioni tendinee nel cavallo mediante Trapianto autologo di cellule mononucleate di midollo osseo (BMMNCS): esperienze cliniche preliminari; proceedings XII Congresso Multisala Società Italiana di Veterinari per Equini (SIVE), Bologna (Italy). January 28. 2006; 162-163
27 Herovici C. A polychrome stain for differentiating pre-collagen from collagen.. Stain Tech 1963; 38: 304-305.
28 Whittaker P, Canham PB. Demonstration of quantitative fabric analysis of tendon collagen using two-dimensional polarized light microscopy.. Matrix 1991; 11: 56-62.
29 Sodersten F, Ekman S, Eloranta ML. et al. Ultrastructural immunolocalization of cartilage oligomeric matrix protein (COMP) in relation to collagen fibrils in the equine tendon.. Matrix Biol 2005; 24: 376-385.
30 Eriksen HA, Pajala A, Leppilahti J. et al. Increased content of type III collagen at the rupture site of human Achilles tendon.. J Orthop Res 2002; 20: 1352-1357.
31 Bosch P, Hertz H, Lintner F. et al. Does the fibrin glue accelerate the healing of tendons? [Transkation].. Arch Orthop Trauma Surg 1981; 98: 305-310.
32 Thomopoulos S, Soslowsky LJ, Flanagan CL. et al. The effect of fibrin clot on healing rat supraspinatus tendon defects.. J Shoulder Elbow Surg 2002; 11: 239-247.
33 Thermann H, Frerichs O, Holch M. et al. Healing of Achilles tendon, an experimental study: part 2--Histological, immunohistological and ultra sonographic analysis.. Foot Ankle Int 2002; 23: 606-613.
34 Simmons PJ, Torok-Storb B. CD34 expression by stromal precursors in normal human adult bone marrow.. Blood 1991; 78: 2848-2853.