Compression Bioreactor-Based Mechanical Loading Induces Mobilization of Human Bone Marrow-Derived Mesenchymal Stromal Cells into Collagen Scaffolds In Vitro

Articular cartilage (AC) is an avascular tissue composed of scattered chondrocytes embedded in a dense extracellular matrix, in which nourishment takes place via the synovial fluid at the surface. AC has a limited intrinsic healing capacity, and thus mainly surgical techniques have been used to relieve pain and improve function. Approaches to promote regeneration remain challenging. The microfracture (MF) approach targets the bone marrow (BM) as a source of factors and progenitor cells to heal chondral defects in situ by opening small holes in the subchondral bone. However, the original function of AC is not obtained yet. We hypothesize that mechanical stimulation can mobilize mesenchymal stromal cells (MSCs) from BM reservoirs upon MF of the subchondral bone. Thus, the aim of this study was to compare the counts of mobilized human BM-MSCs (hBM-MSCs) in alginate-laminin (alginate-Ln) or collagen-I (col-I) scaffolds upon intermittent mechanical loading. The mechanical set up within an established bioreactor consisted of 10% strain, 0.3 Hz, breaks of 10 s every 180 cycles for 24 h. Contrary to previous findings using porcine MSCs, no significant cell count was found for hBM-MSCs into alginate-Ln scaffolds upon mechanical stimulation (8 ± 5 viable cells/mm3 for loaded and 4 ± 2 viable cells/mm3 for unloaded alginate-Ln scaffolds). However, intermittent mechanical stimulation induced the mobilization of hBM-MSCs into col-I scaffolds 10-fold compared to the unloaded col-I controls (245 ± 42 viable cells/mm3 vs. 22 ± 6 viable cells/mm3, respectively; p-value < 0.0001). Cells that mobilized into the scaffolds by mechanical loading did not show morphological changes. This study confirmed that hBM-MSCs can be mobilized in vitro from a reservoir toward col-I but not alginate-Ln scaffolds upon intermittent mechanical loading, against gravity.

[1]  E. Paluch,et al.  Of Cell Shapes and Motion: The Physical Basis of Animal Cell Migration. , 2020, Developmental cell.

[2]  N. Gretz,et al.  Bioreactor for mobilization of mesenchymal stem/stromal cells into scaffolds under mechanical stimulation: Preliminary results , 2020, PloS one.

[3]  M. Stoffel,et al.  Chondrocyte colonisation of a tissue-engineered cartilage substitute under a mechanical stimulus. , 2019, Medical engineering & physics.

[4]  C. Ahn,et al.  Simulated microgravity with floating environment promotes migration of non-small cell lung cancers , 2019, Scientific Reports.

[5]  C. Voermans,et al.  Nuclear shape, protrusive behaviour and in vivo retention of human bone marrow mesenchymal stromal cells is controlled by Lamin-A/C expression , 2019, Scientific Reports.

[6]  Michael Sixt,et al.  Mechanisms of 3D cell migration , 2019, Nature Reviews Molecular Cell Biology.

[7]  Freddie H. Fu,et al.  Dynamic Compressive Loading Improves Cartilage Repair in an In Vitro Model of Microfracture: Comparison of 2 Mechanical Loading Regimens on Simulated Microfracture Based on Fibrin Gel Scaffolds Encapsulating Connective Tissue Progenitor Cells , 2019, The American journal of sports medicine.

[8]  S. Marlovits,et al.  [Significance of Matrix-augmented Bone Marrow Stimulation for Treatment of Cartilage Defects of the Knee: A Consensus Statement of the DGOU Working Group on Tissue Regeneration]. , 2018, Zeitschrift fur Orthopadie und Unfallchirurgie.

[9]  A. Higuchi,et al.  Collagen Scaffolds in Cartilage Tissue Engineering and Relevant Approaches for Future Development , 2018, Tissue Engineering and Regenerative Medicine.

[10]  F. MacKintosh,et al.  The Role of Network Architecture in Collagen Mechanics. , 2018, Biophysical journal.

[11]  Jane Ru Choi,et al.  Effects of mechanical loading on human mesenchymal stem cells for cartilage tissue engineering , 2018, Journal of cellular physiology.

[12]  N. Benkirane-Jessel,et al.  Mechanical stimulations on human bone marrow mesenchymal stem cells enhance cells differentiation in a three‐dimensional layered scaffold , 2018, Journal of tissue engineering and regenerative medicine.

[13]  C. Lim,et al.  Microfluidic label-free selection of mesenchymal stem cell subpopulation during culture expansion extends the chondrogenic potential in vitro. , 2018, Lab on a chip.

[14]  M. Alini,et al.  Mechanical stimulation of mesenchymal stem cells: Implications for cartilage tissue engineering , 2017, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[15]  Gopal Shankar Krishnakumar,et al.  Evaluation of different crosslinking agents on hybrid biomimetic collagen-hydroxyapatite composites for regenerative medicine. , 2018, International journal of biological macromolecules.

[16]  Devon E. Anderson,et al.  Dynamic Mechanical Compression of Chondrocytes for Tissue Engineering: A Critical Review , 2017, Front. Bioeng. Biotechnol..

[17]  David R. Christian,et al.  Arthroscopic Management of Isolated Tibial Plateau Defect With Microfracture and Micronized Allogeneic Cartilage–Platelet-Rich Plasma Adjunct , 2017, Arthroscopy techniques.

[18]  M. Marcacci,et al.  Investigation of different cross-linking approaches on 3D gelatin scaffolds for tissue engineering application: A comparative analysis. , 2017, International journal of biological macromolecules.

[19]  T. Hewett,et al.  Microfracture of Articular Cartilage , 2016, JBJS reviews.

[20]  U. Schneider Controlled, randomized multicenter study to compare compatibility and safety of ChondroFiller liquid (cell free 2-component collagen gel) with microfracturing of patients with focal cartilage defects of the knee joint , 2016 .

[21]  G. Im Endogenous Cartilage Repair by Recruitment of Stem Cells. , 2016, Tissue engineering. Part B, Reviews.

[22]  X Zhang,et al.  Fate decision of mesenchymal stem cells: adipocytes or osteoblasts? , 2016, Cell Death and Differentiation.

[23]  J. Jerosch,et al.  Retrospektive Untersuchung einer zellfreien Matrix zur Knorpeltherapie , 2016 .

[24]  S. Grässel Collagens in Hyaline Cartilage , 2016 .

[25]  Anja Nohe,et al.  Role of Chondrocytes in Cartilage Formation, Progression of Osteoarthritis and Cartilage Regeneration , 2015, Journal of developmental biology.

[26]  Youngmee Jung,et al.  In Situ Recruitment of Human Bone Marrow-Derived Mesenchymal Stem Cells Using Chemokines for Articular Cartilage Regeneration , 2015, Cell transplantation.

[27]  S. Mousavi,et al.  Three-Dimensional Numerical Model of Cell Morphology during Migration in Multi-Signaling Substrates , 2015, PloS one.

[28]  Jinxi Wang,et al.  Cell-based articular cartilage repair: the link between development and regeneration. , 2015, Osteoarthritis and cartilage.

[29]  K. Schüttler,et al.  Short-term follow up after implantation of a cell-free collagen type I matrix for the treatment of large cartilage defects of the knee , 2015, International Orthopaedics.

[30]  C. Madeira,et al.  Advanced cell therapies for articular cartilage regeneration. , 2015, Trends in biotechnology.

[31]  Jerry C. Hu,et al.  Repair and tissue engineering techniques for articular cartilage , 2015, Nature Reviews Rheumatology.

[32]  Guillaume Charras,et al.  Physical influences of the extracellular environment on cell migration , 2014, Nature Reviews Molecular Cell Biology.

[33]  K. Tryggvason,et al.  Monolayer culturing and cloning of human pluripotent stem cells on laminin-521–based matrices under xeno-free and chemically defined conditions , 2014, Nature Protocols.

[34]  Huijun Sun,et al.  Icariin promotes directed chondrogenic differentiation of bone marrow mesenchymal stem cells but not hypertrophy in vitro , 2014, Experimental and therapeutic medicine.

[35]  Farshid Guilak,et al.  The Mechanobiology of Articular Cartilage: Bearing the Burden of Osteoarthritis , 2014, Current Rheumatology Reports.

[36]  M. H. Doweidar,et al.  A novel mechanotactic 3D modeling of cell morphology , 2014, Physical biology.

[37]  J. Kere,et al.  Clonal culturing of human embryonic stem cells on laminin-521/E-cadherin matrix in defined and xeno-free environment , 2014, Nature Communications.

[38]  Li Duan,et al.  Extracellular matrix production in vitro in cartilage tissue engineering , 2014, Journal of Translational Medicine.

[39]  M. Mercado,et al.  Cross-Linked Alginate Film Pore Size Determination Using Atomic Force Microscopy and Validation Using Diffusivity Determinations * , 2013 .

[40]  S. Paliwal,et al.  A Review of the Metabolism of 1,4-Butanediol Diglycidyl Ether–Crosslinked Hyaluronic Acid Dermal Fillers , 2013, Dermatologic surgery : official publication for American Society for Dermatologic Surgery [et al.].

[41]  Farshid Guilak,et al.  Diurnal variations in articular cartilage thickness and strain in the human knee. , 2013, Journal of biomechanics.

[42]  P. Doevendans,et al.  Human versus porcine mesenchymal stromal cells: phenotype, differentiation potential, immunomodulation and cardiac improvement after transplantation , 2012, Journal of cellular and molecular medicine.

[43]  M. Knight,et al.  Dynamic compressive strain influences chondrogenic gene expression in human periosteal cells: a case study. , 2012, Journal of the mechanical behavior of biomedical materials.

[44]  D. Hutmacher,et al.  CD73 and CD29 concurrently mediate the mechanically induced decrease of migratory capacity of mesenchymal stromal cells. , 2011, European cells & materials.

[45]  P. Neyret,et al.  MACI - a new era? , 2011, Sports medicine, arthroscopy, rehabilitation, therapy & technology : SMARTT.

[46]  W. Hanke,et al.  Effects of Altered Gravity on the Actin and Microtubule Cytoskeleton, Cell Migration and Neurite Outgrowth , 2011 .

[47]  J. Wang Cell Traction Forces (CTFs) and CTF Microscopy Applications in Musculoskeletal Research. , 2010, Operative techniques in orthopaedics.

[48]  S. Stolnik,et al.  Alginate encapsulation technology supports embryonic stem cells differentiation into insulin-producing cells. , 2009, Journal of biotechnology.

[49]  W. Richter,et al.  Mesenchymal stem cells and cartilage in situ regeneration , 2009, Journal of internal medicine.

[50]  M. Endres,et al.  Formation of cartilage repair tissue in articular cartilage defects pretreated with microfracture and covered with cell‐free polymer‐based implants , 2009, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[51]  Robert A. Brown,et al.  Guiding cell migration in 3D: a collagen matrix with graded directional stiffness. , 2009, Cell motility and the cytoskeleton.

[52]  J. Steinmeyer,et al.  Intermittent mechanical loading of articular cartilage explants modulates chondroitin sulfate fine structure. , 2007, Osteoarthritis and cartilage.

[53]  Moonsoo Jin,et al.  Shear and Compression Differentially Regulate Clusters of Functionally Related Temporal Transcription Patterns in Cartilage Tissue* , 2006, Journal of Biological Chemistry.

[54]  Hermann Eichler,et al.  Comparative Analysis of Mesenchymal Stem Cells from Bone Marrow, Umbilical Cord Blood, or Adipose Tissue , 2006, Stem cells.

[55]  G. Duda,et al.  Simulation of cell differentiation in fracture healing: mechanically loaded composite scaffolds in a novel bioreactor system. , 2006, Tissue engineering.

[56]  P. Behrens,et al.  In vivo matrix-guided human mesenchymal stem cells , 2006, Cellular and Molecular Life Sciences.

[57]  J. Steinmeyer,et al.  Proteoglycan metabolism and viability of articular cartilage explants as modulated by the frequency of intermittent loading. , 2003, Osteoarthritis and cartilage.

[58]  J. Feijen,et al.  The kinetics of 1,4-butanediol diglycidyl ether crosslinking of dermal sheep collagen. , 2000, Journal of biomedical materials research.

[59]  A. Grodzinsky,et al.  Cartilage tissue remodeling in response to mechanical forces. , 2000, Annual review of biomedical engineering.

[60]  J. Steinmeyer,et al.  Effects of intermittently applied cyclic loading on proteoglycan metabolism and swelling behaviour of articular cartilage explants. , 1999, Osteoarthritis and cartilage.

[61]  P. Cochard,et al.  Dual staining assessment of Schwann cell viability within whole peripheral nerves using calcein-AM and ethidium homodimer , 1997, Journal of Neuroscience Methods.

[62]  Z. Šidák Rectangular Confidence Regions for the Means of Multivariate Normal Distributions , 1967 .