Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress 2023 EHA Congress 2023 ASCO Annual Meeting
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
International Orthopaedics
Published in: International Orthopaedics 7/2016

01-07-2016 | Original Paper

Mesenchymal stem cells reside in anterior cruciate ligament remnants in situ

Authors: Weili Fu, Qi Li, Xin Tang, Gang Chen, Chenghao Zhang, Jian Li

Published in: International Orthopaedics | Issue 7/2016

Login to get access

Abstract

Purpose

It has been reported that the anterior cruciate ligament (ACL) has certain self-healing ability after acute injury or with primary suture repair. Many studies have confirmed that a remnant preservation technique with ACL reconstruction contributes to biological augmentation for ACL healing. However, it remains unclear whether mesenchymal stem cells (MSC) reside in ACL remnants in situ. The aim of this study was to investigate the methods of culture and identification of MSC derived from the remnants of ACL rupture patients and to analyse these MSC’s properties.

Methods

The cells of ACL remnants from the ACL rupture patients were isolated by the methods of enzymatic digestion and cultured in vitro to the third passage under the microscope to observe their morphology and growth status. The third passage of isolated cells was analysed for the identification of immunophenotype, osteogenic, adipogenic and chondrogenic differentiation.

Results

On the third to fifth days of in vitro culture, a few cells of long fusiform shape appeared and were adherent to the plastic walls. On the sixth to ninth days, cells clustered and colonies were observed. The third passage cells showed uniform cell morphology and good proliferation, with appearance of the typical surface markers of MSC, CD29, CD44, CD90 and CD105. The surface markers of CD34 and CD45 of haematopoietic stem cells were not expressed. Under appropriate conditions of in vitro culture, isolated cells could be differentiated into osteoblasts that deposit mineralised matrix and express early osteogenic markers, adipocytes that accumulate lipid droplets in cytoplasm and chondrocytes that secrete chondrogenic-specific matrix aggrecan and collagen II. Real-time polymerase chain reaction (PCR) analysis demonstrated that the specific mRNA expression of osteogenesis, adipogenesis and chondrogenesis increased significantly compared with the control groups at day zero.

Conclusions

Stem cells derived in situ from the human ACL stump were successfully isolated and characterised. Those isolated cells were identified as MSC according to their adherent ability, morphology, surface markers and multilineage differentiation potential. MSC derived from ACL remnants could be a potential source of seeding cells for ligament regeneration.
Literature
1.
Stevanović V, Blagojević Z, Petković A, Glišić M, Sopta J, Nikolić V, Milisavljević M (2013) Semitendinosus tendon regeneration after anterior cruciate ligament reconstruction: can we use it twice? Int Orthop 37:2475–2481CrossRefPubMedPubMedCentral
2.
Sun L, Wu B, Tian M, Liu B, Luo Y (2013) Comparison of graft healing in anterior cruciate ligament reconstruction with and without a preserved remnant in rabbits. Knee 20:537–544CrossRefPubMed
3.
4.
Oiestad BE, Holm I, Aune AK, Gunderson R, Myklebust G, Engebretsen L, Fosdahl MA, Risberg M (2010) Knee function and prevalence of knee osteoarthritis after anterior cruciate ligament reconstruction: a prospective study with 10 to 15 years of follow-up. Am J Sports Med 38:2201–2210CrossRefPubMed
5.
6.
Schwarting T, Benölken M, Ruchholtz S, Frink M, Lechler P (2015) Bone morphogenetic protein-7 enhances bone-tendon integration in a murine in vitro co-culture model. Int Orthop 39:799–805CrossRefPubMed
7.
Struewer J, Roessler PP, Schuettler KF, Ruppert V, Stein T, Timmesfeld N, Paletta JR, Efe T (2014) Influence of cyclical mechanical loading on osteogenic markers in an osteoblast-fibroblast co-culture in vitro: tendon-to-bone interface in anterior cruciate ligament reconstruction. Int Orthop 38:1083–1089CrossRefPubMed
8.
Leong NL, Petrigliano FA, McAllister DR (2014) Current tissue engineering strategies in anterior cruciate ligament reconstruction. J Biomed Mater Res A 102:1614–1624CrossRefPubMed
9.
Yang G, Rothrauff BB, Tuan RS (2013) Tendon and ligament regeneration and repair: clinical relevance and developmental paradigm. Birth Defects Res C Embryo Today 99:203–222CrossRefPubMedPubMedCentral
10.
Park SY, Oh H, Park SW, Lee JH, Lee SH, Yoon KH (2012) Clinical outcomes of remnant-preserving augmentation versus double-bundle reconstruction in the anterior cruciate ligament reconstruction. Arthroscopy 28:1833–1841CrossRefPubMed
11.
Kazusa H, Nakamae A, Ochi M (2013) Augmentation technique for anterior cruciate ligament injury. Clin Sports Med 32:127–140CrossRefPubMed
12.
Wu B, Zhao Z, Li S, Sun L (2013) Preservation of remnant attachment improves graft healing in a rabbit model of anterior cruciate ligament reconstruction. Arthroscopy 29:1362–1371CrossRefPubMed
13.
Eguchi A, Adachi N, Nakamae A, Usman MA, Deie M, Ochi M (2014) Proprioceptive function after isolated single-bundle posterior cruciate ligament reconstruction with remnant preservation for chronic posterior cruciate ligament injuries. Orthop Traumatol Surg Res 100:303–308CrossRefPubMed
14.
Lee DC, Shon OJ, Kwack BH, Lee SJ (2013) Proprioception and clinical results of anterolateral single-bundle posterior cruciate ligament reconstruction with remnant preservation. Knee Surg Relat Res 25:126–132CrossRefPubMedPubMedCentral
15.
Kim SJ, Kim SH, Chun YM, Hwang BY, Choi DH, Yoon JY (2012) Clinical comparison of conventional and remnant-preserving transtibial single-bundle posterior cruciate ligament reconstruction combined with posterolateral corner reconstruction. Am J Sports Med 40:640–649CrossRefPubMed
16.
17.
Lui PP, Chan KM (2011) Tendon-derived stem cells (TDSCs): from basic science to potential roles in tendon pathology and tissue engineering applications. Stem Cell Rev 7:883–897CrossRefPubMed
18.
Fu WL, Zhou CY, Yu JK (2014) A new source of mesenchymal stem cells for articular cartilage repair: MSCs derived from mobilized peripheral blood share similar biological characteristics in vitro and chondrogenesis in vivo as MSCs from bone marrow in a rabbit model. Am J Sports Med 42:592–601CrossRefPubMed
19.
Fu WL, Zhang JY, Fu X, Duan XN, Leung KK, Jia ZQ, Wang WP, Zhou CY, Yu JK (2012) Comparative study of the biological characteristics of mesenchymal stem cells from bone marrow and peripheral blood of rats. Tissue Eng Part A 18:1793–1803CrossRefPubMed
20.
Segawa Y, Muneta T, Makino H, Nimura A, Mochizuki T, Ju YJ, Ezura Y, Umezawa A, Sekiya I (2009) Mesenchymal stem cells derived from synovium, meniscus, anterior cruciate ligament, and articular chondrocytes share similar gene expression profiles. J Orthop Res 27:435–441CrossRefPubMed
21.
Asari T, Furukawa K, Tanaka S, Kudo H, Mizukami H, Ono A, Numasawa T, Kumagai G, Motomura S, Yagihashi S, Toh S (2012) Mesenchymal stem cell isolation and characterization from human spinal ligaments. Biochem Biophys Res Commun 417:1193–1199CrossRefPubMed
22.
Crisan M, Yap S, Casteilla L, Chen CW, Corselli M, Park TS, Andriolo G, Sun B, Zheng B, Zhang L, Norotte C, Teng PN, Traas J, Schugar R, Deasy BM, Badylak S, Buhring HJ, Giacobino JP, Lazzari L, Huard J, Péault B (2008) A perivascular origin for mesenchymal stem cells in multiple human organs. Cell Stem Cell 3:301–313CrossRefPubMed
23.
da Silva Meirelles L, Caplan AI, Nardi NB (2008) In search of the in vivo identity of mesenchymal stem cells. Stem Cells 26:2287–2299CrossRefPubMed
24.
25.
Mifune Y, Matsumoto T, Ota S, Nishimori M, Usas A, Kopf S, Kuroda R, Kurosaka M, Fu FH, Huard J (2012) Therapeutic potential of anterior cruciate ligament-derived stem cells for anterior cruciate ligament reconstruction. Cell Transplant 21:1651–1665CrossRefPubMed
26.
Nohmi S, Yamamoto Y, Mizukami H, Ishibashi Y, Tsuda E, Maniwa K, Yagihashi S, Motomura S, Toh S, Furukawa K (2012) Post injury changes in the properties of mesenchymal stem cells derived from human anterior cruciate ligaments. Int Orthop 36:1515–1522CrossRefPubMedPubMedCentral
27.
Kwapisz A, Chojnacki M, Domzalski M, Grzegorzewski A, Synder M (2014) Do gene expression changes in articular cartilage proteases of the synovial membrane correlate with expression changes of the same genes in systemic blood cells? Int Orthop 38:649–654CrossRefPubMed
28.
Morito T, Muneta T, Hara K, Ju YJ, Mochizuki T, Makino H, Umezawa A, Sekiya I (2008) Synovial fluid-derived mesenchymal stem cells increase after intra-articular ligament injury in humans. Rheumatology (Oxford) 47:1137–1143CrossRef
29.
Zhang J, Pan T, Im HJ, Fu FH, Wang JH (2011) Differential properties of human ACL and MCL stem cells may be responsible for their differential healing capacity. BMC Med 9:68CrossRefPubMedPubMedCentral
30.
31.
Bi Y, Ehirchiou D, Kilts TM, Inkson CA, Embree MC, Sonoyama W, Li L, Leet AI, Seo BM, Zhang L, Shi S, Young MF (2007) Identification of tendon stem/progenitor cells and the role of the extracellular matrix in their niche. Nat Med 13:1219–1227CrossRefPubMed
32.
Pauly S, Klatte F, Strobel C, Schmidmaier G, Greiner S, Scheibel M, Wildemann B (2010) Characterization of tendon cell cultures of the human rotator cuff. Eur Cell Mater 20:84–97PubMed
33.
Van Eijk F, Saris DB, Riesle J, Willems WJ, Van Blitterswijk CA, Verbout AJ, Dhert WJ (2004) Tissue engineering of ligaments: a comparison of bone marrow stromal cells, anterior cruciate ligament, and skin fibroblasts as cell source. Tissue Eng 10:893–903CrossRefPubMed
34.
Cheng MT, Yang HW, Chen TH, Lee OK (2009) Isolation and characterization of multipotent stem cells from human cruciate ligaments. Cell Prolif 42:448–460CrossRefPubMed
35.
Cheng MT, Liu CL, Chen TH, Lee OK (2010) Comparison of potentials between stem cells isolated from human anterior cruciate ligament and bone marrow for ligament tissue engineering. Tissue Eng Part A 16:2237–2253CrossRefPubMed
36.
Randelli P, Conforti E, Piccoli M, Ragone V, Creo P, Cirillo F, Masuzzo P, Tringali C, Cabitza P, Tettamanti G, Gagliano N, Anastasia L (2013) Isolation and characterization of 2 new human rotator cuff and long head of biceps tendon cells possessing stem cell-like self-renewal and multipotential differentiation capacity. Am J Sports Med 41:1653–1664CrossRefPubMed
Metadata
Title
Mesenchymal stem cells reside in anterior cruciate ligament remnants in situ
Authors
Weili Fu
Qi Li
Xin Tang
Gang Chen
Chenghao Zhang
Jian Li
Publication date
01-07-2016
Publisher
Springer Berlin Heidelberg
Published in
International Orthopaedics / Issue 7/2016
Print ISSN: 0341-2695
Electronic ISSN: 1432-5195
DOI
https://doi.org/10.1007/s00264-015-2925-1

Other articles of this Issue 7/2016

International Orthopaedics 7/2016 Go to the issue

Original Paper

Influence of somatization and depressive symptoms on the course of pain within the first year after uncomplicated total knee replacement: a prospective study

Original Paper

Treatment of feet deformities in epidermolysis bullosa

Original Paper

Long-term follow-up of primary total hip arthroplasty with the Alloclassic Variall system

Original Paper

A prospective cohort study of the clinical presentation of non-traumatic osteonecrosis of the femoral head: spine and knee symptoms as clinical presentation of hip osteonecrosis

Original Paper

Polymicrobial infections reduce the cure rate in prosthetic joint infections: outcome analysis with two-stage exchange and follow-up ≥two years

Editorial

Complications and rare cases in orthopaedics