Collagen‐containing scaffolds enhance attachment and proliferation of non‐cultured bone marrow multipotential stromal cells

Large bone defects are ideally treated with autografts, which have many limitations. Therefore, osteoconductive scaffolds loaded with autologous bone marrow (BM) aspirate are increasingly used as alternatives. The purpose of this study was to compare the growth of multipotential stromal cells (MSCs) from unprocessed BM on a collagen‐containing bovine bone scaffold (Orthoss® Collagen) with a non‐collagen‐containing bovine bone scaffold, Orthoss®. Another collagen‐containing synthetic scaffold, Vitoss® was included in the comparison. Colonization of scaffolds by BM MSCs (n = 23 donors) was evaluated using microscopy, colony forming unit‐fibroblast assay and flow‐cytometry. The number of BM MSCs initially attached to Orthoss® Collagen and Vitoss® was similar but greater than Orthoss® (p = 0.001 and p = 0.041, respectively). Furthermore, the number of MSCs released from Orthoss® Collagen and Vitoss® after 2‐week culture was also higher compared to Orthoss® (p = 0.010 and p = 0.023, respectively). Interestingly, collagen‐containing scaffolds accommodated larger numbers of lymphocytic and myelomonocytic cells. Additionally, the proliferation of culture‐expanded MSCs on Orthoss® collagen and Vitoss® was greater compared to Orthoss® (p = 0.047 and p = 0.004, respectively). Collectively, collagen‐containing scaffolds were superior in supporting the attachment and proliferation of MSCs when they were loaded with unprocessed BM aspirates. This highlights the benefit of collagen incorporation into bone scaffolds for use with autologous bone marrow aspirates as autograft substitutes. © 2015 The Authors. Journal of Orthopaedic Research Published by Wiley Periodicals, Inc. on behalf of Orthopaedic Research Society. J Orthop Res 34:597–606, 2016.

[1]  TorreMaria Luisa,et al.  Ex Vivo Expanded Mesenchymal Stromal Cell Minimal Quality Requirements for Clinical Application , 2015 .

[2]  P. Layrolle,et al.  Osteoblastic and osteoclastic differentiation of human mesenchymal stem cells and monocytes in a miniaturized three-dimensional culture with mineral granules. , 2014, Acta biomaterialia.

[3]  G. Calori,et al.  Incidence of donor site morbidity following harvesting from iliac crest or RIA graft. , 2014, Injury.

[4]  P. Giannoudis,et al.  Multipotential stromal cell abundance in cellular bone allograft: comparison with fresh age-matched iliac crest bone and bone marrow aspirate. , 2014, Regenerative medicine.

[5]  J. M. Guimarães,et al.  The effect of autologous concentrated bone-marrow grafting on the healing of femoral shaft non-unions after locked intramedullary nailing. , 2014, Injury.

[6]  G. Logroscino,et al.  Bone substitutes in orthopaedic surgery: from basic science to clinical practice , 2014, Journal of Materials Science: Materials in Medicine.

[7]  Nicola Maffulli,et al.  Bone regenerative medicine: classic options, novel strategies, and future directions , 2014, Journal of Orthopaedic Surgery and Research.

[8]  P. Giannoudis,et al.  Native multipotential stromal cell colonization and graft expander potential of a bovine natural bone scaffold , 2013, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[9]  V. Kuzmenko,et al.  Universal method for protein bioconjugation with nanocellulose scaffolds for increased cell adhesion. , 2013, Materials science & engineering. C, Materials for biological applications.

[10]  M. Anders,et al.  Donor site morbidity with reamer-irrigator-aspirator (RIA) use for autogenous bone graft harvesting in a single centre 204 case series. , 2013, Injury.

[11]  P. Hernigou,et al.  Benefits of small volume and small syringe for bone marrow aspirations of mesenchymal stem cells , 2013, International Orthopaedics.

[12]  W. Khan,et al.  A systematic review on preclinical and clinical studies on the use of scaffolds for bone repair in skeletal defects. , 2013, Current stem cell research & therapy.

[13]  G. Duda,et al.  Crosstalk between immune cells and mesenchymal stromal cells in a 3D bioreactor system. , 2012, The International journal of artificial organs.

[14]  T. Ma,et al.  Regulation of autocrine fibroblast growth factor‐2 signaling by perfusion flow in 3D human mesenchymal stem cell constructs , 2012, Biotechnology progress.

[15]  J. Ge,et al.  Serum Starvation Induced Cell Cycle Synchronization Facilitates Human Somatic Cells Reprogramming , 2012, PloS one.

[16]  P. Giannoudis,et al.  Single-platform quality control assay to quantify multipotential stromal cells in bone marrow aspirates prior to bulk manufacture or direct therapeutic use. , 2012, Cytotherapy.

[17]  P. Giannoudis,et al.  High abundance of CD271+ multipotential stromal cells (MSCs) in intramedullary cavities of long bones , 2012, Bone.

[18]  P. Genever,et al.  Effects of endothelial cells on human mesenchymal stem cell activity in a three-dimensional in vitro model. , 2011, European cells & materials.

[19]  A. Polini,et al.  Osteoinduction of Human Mesenchymal Stem Cells by Bioactive Composite Scaffolds without Supplemental Osteogenic Growth Factors , 2011, PloS one.

[20]  Gerhard Schmidmaier,et al.  What should be the characteristics of the ideal bone graft substitute? Combining scaffolds with growth factors and/or stem cells. , 2011, Injury.

[21]  Liesbet Geris,et al.  The combined bone forming capacity of human periosteal derived cells and calcium phosphates. , 2011, Biomaterials.

[22]  D. Docheva,et al.  Integrins α2β1 and α11β1 regulate the survival of mesenchymal stem cells on collagen I , 2011, Cell Death and Disease.

[23]  Xuebin B. Yang,et al.  Mesenchymal stem cells and bone regeneration: current status. , 2011, Injury.

[24]  Baoqiang Li,et al.  Improving bone marrow stromal cell attachment on chitosan/hydroxyapatite scaffolds by an immobilized RGD peptide , 2010, Biomedical materials.

[25]  R. Reis,et al.  Enzymatic degradation of 3D scaffolds of starch-poly-(ɛ-caprolactone) prepared by supercritical fluid technology , 2010 .

[26]  R. Windhager,et al.  Behaviour of multipotent maxillary bone-derived cells on beta-tricalcium phosphate and highly porous bovine bone mineral. , 2010, Clinical oral implants research.

[27]  P. Emery,et al.  Large-scale extraction and characterization of CD271+ multipotential stromal cells from trabecular bone in health and osteoarthritis: implications for bone regeneration strategies based on uncultured or minimally cultured multipotential stromal cells. , 2010, Arthritis and rheumatism.

[28]  D. Griffon,et al.  Effect of collagen II coating on mesenchymal stem cell adhesion on chitosan and on reacetylated chitosan fibrous scaffolds , 2010, Journal of materials science. Materials in medicine.

[29]  Xiao-Dong Chen Extracellular matrix provides an optimal niche for the maintenance and propagation of mesenchymal stem cells. , 2010, Birth defects research. Part C, Embryo today : reviews.

[30]  Adam J Engler,et al.  Intrinsic extracellular matrix properties regulate stem cell differentiation. , 2010, Journal of biomechanics.

[31]  Yilin Cao,et al.  Repair of goat tibial defects with bone marrow stromal cells and β-tricalcium phosphate , 2008, Journal of materials science. Materials in medicine.

[32]  E. Jones,et al.  Age-related changes in human bone marrow-derived mesenchymal stem cells: Consequences for cell therapies , 2008, Mechanisms of Ageing and Development.

[33]  N. Boiret-Dupré,et al.  Mesenchymal content of fresh bone marrow: a proposed quality control method for cell therapy , 2007, British journal of haematology.

[34]  Maurilio Marcacci,et al.  Stem cells associated with macroporous bioceramics for long bone repair: 6- to 7-year outcome of a pilot clinical study. , 2007, Tissue engineering.

[35]  K. Koval,et al.  Bone grafts and bone graft substitutes in orthopaedic trauma surgery. A critical analysis. , 2007, The Journal of bone and joint surgery. American volume.

[36]  Eleftherios Tsiridis,et al.  Bone substitutes: an update. , 2005, Injury.

[37]  Xuebin B. Yang,et al.  Biomimetic collagen scaffolds for human bone cell growth and differentiation. , 2004, Tissue engineering.

[38]  G. Kopen,et al.  Characterization of mesenchymal stem cells isolated from murine bone marrow by negative selection , 2003, Journal of cellular biochemistry.

[39]  V. Goldberg,et al.  The Effect of Implants Loaded with Autologous Mesenchymal Stem Cells on the Healing of Canine Segmental Bone Defects* , 1998, The Journal of bone and joint surgery. American volume.

[40]  G. Alessandri,et al.  Ex vivo expanded mesenchymal stromal cell minimal quality requirements for clinical application. , 2015, Stem cells and development.

[41]  Philippe Rosset,et al.  Bone fracture healing: cell therapy in delayed unions and nonunions. , 2015, Bone.

[42]  M. Blanco-Gelaz,et al.  Quantifying mesenchymal stem cells in the mononuclear cell fraction of bone marrow samples obtained for cell therapy. , 2013, Transplantation proceedings.

[43]  J. Lindhe,et al.  Dynamics of Bio-Oss Collagen incorporation in fresh extraction wounds: an experimental study in the dog. , 2010, Clinical oral implants research.

[44]  S. Deb,et al.  In vitro proliferation and differentiation of human mesenchymal stem cells on hydroxyapatite versus human demineralised bone matrix with and without osteogenic protein-1. , 2009, Expert opinion on biological therapy.

[45]  M. Pittenger,et al.  Mesenchymal stem cells from adult bone marrow. , 2008, Methods in molecular biology.

[46]  K. Pattanapanyasat,et al.  New BD FACSCount™ CD4 reagent system for simultaneous enumeration of percent and absolute CD4 T‐lymphocytes in HIV‐1‐infected pediatric patients , 2008, Cytometry. Part B, Clinical cytometry.

[47]  D. Prockop,et al.  Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. , 2006, Cytotherapy.