A new platelet cryoprecipitate glue promoting bone formation after ectopic mesenchymal stromal cell-loaded biomaterial implantation in nude mice

IntroductionThis study investigated the promising effect of a new Platelet Glue obtained from Cryoprecipitation of Apheresis Platelet products (PGCAP) used in combination with Mesenchymal Stromal Cells (MSC) loaded on ceramic biomaterials to provide novel strategies enhancing bone repair.MethodsPGCAP growth factor content was analyzed by ELISA and compared to other platelet and plasma-derived products. MSC loaded on biomaterials (65% hydroxyapatite/35% beta-TCP or 100% beta-TCP) were embedded in PGCAP and grown in presence or not of osteogenic induction medium for 21 days. Biomaterials were then implanted subcutaneously in immunodeficient mice for 28 days. Effect of PGCAP on MSC was evaluated in vitro by proliferation and osteoblastic gene expression analysis and in vivo by histology and immunohistochemistry.ResultsWe showed that PGCAP, compared to other platelet-derived products, allowed concentrating large amount of growth factors and cytokines which promoted MSC and osteoprogenitor proliferation. Next, we found that PGCAP improves the proliferation of MSC and osteogenic-induced MSC. Furthermore, we demonstrated that PGCAP up-regulates the mRNA expression of osteogenic markers (Collagen type I, Osteonectin, Osteopontin and Runx2). In vivo, type I collagen expressed in ectopic bone-like tissue was highly enhanced in biomaterials embedded in PGCAP in the absence of osteogenic pre-induction. Better results were obtained with 65% hydroxyapatite/35% beta-TCP biomaterials as compared to 100% beta-TCP.ConclusionsWe have demonstrated that PGCAP is able to enhance in vitro MSC proliferation, osteoblastic differentiation and in vivo bone formation in the absence of osteogenic pre-induction. This clinically adaptable platelet glue could be of interest for improving bone repair.

[1]  H. Klüter,et al.  Autologous concentrated platelet-rich plasma (cPRP) for local application in bone regeneration. , 2002, International journal of oral and maxillofacial surgery.

[2]  Huipin Yuan,et al.  Osteoinductive ceramics as a synthetic alternative to autologous bone grafting , 2010, Proceedings of the National Academy of Sciences.

[3]  G. Daculsi,et al.  Influence of biphasic calcium phosphate granulometry on bone ingrowth, ceramic resorption, and inflammatory reactions: preliminary in vitro and in vivo study. , 1999, Journal of biomedical materials research.

[4]  K. Heiple,et al.  Biology of cancellous bone grafts. , 1987, The Orthopedic clinics of North America.

[5]  N. Kulagina,et al.  Fibroblast precursors in normal and irradiated mouse hematopoietic organs. , 1976, Experimental hematology.

[6]  M. Barbeck,et al.  The chemical composition of synthetic bone substitutes influences tissue reactions in vivo: histological and histomorphometrical analysis of the cellular inflammatory response to hydroxyapatite, beta-tricalcium phosphate and biphasic calcium phosphate ceramics , 2012, Biomedical materials.

[7]  Krishnendu Roy,et al.  Biomaterials for stem cell differentiation. , 2008, Advanced drug delivery reviews.

[8]  C. Choong,et al.  MSCs into adipogenic, chondrogenic, and osteogenic lineages identify markers and signaling pathways important in differentiation of growth of mesenchymal stem cells (MSCs): transcriptional profiling can , and FGF signaling is important for differentiation and β PDGF, TGF- , 2013 .

[9]  David L. Kaplan,et al.  BMP-silk composite matrices heal critically sized femoral defects. , 2007, Bone.

[10]  D. Busch,et al.  Platelets secrete stromal cell–derived factor 1α and recruit bone marrow–derived progenitor cells to arterial thrombi in vivo , 2006, The Journal of experimental medicine.

[11]  R. Marx,et al.  Platelet-rich plasma: Growth factor enhancement for bone grafts. , 1998, Oral surgery, oral medicine, oral pathology, oral radiology, and endodontics.

[12]  Ronald E. Unger,et al.  Influence of β-tricalcium phosphate granule size and morphology on tissue reaction in vivo. , 2010, Acta biomaterialia.

[13]  R. Albin.  Regeneration , 1993, Neurology.

[14]  Joshua D. Dilley,et al.  The Use of Near-Infrared Spectroscopy As a Substitute for Blood Pressure Monitoring in a Patient with Severe Osteogenesis Imperfecta , 2012 .

[15]  A. Nguyen,et al.  Brief review of models of ectopic bone formation. , 2012, Stem cells and development.

[16]  A. Uccelli,et al.  Immunoregulatory function of mesenchymal stem cells , 2006, European journal of immunology.

[17]  A Boyde,et al.  Autologous bone marrow stromal cells loaded onto porous hydroxyapatite ceramic accelerate bone repair in critical-size defects of sheep long bones. , 2000, Journal of biomedical materials research.

[18]  T. Hansen,et al.  Effect of platelet concentration in platelet-rich plasma on peri-implant bone regeneration. , 2004, Bone.

[19]  K. Karkouti,et al.  Cryoprecipitate: the current state of knowledge. , 2009, Transfusion medicine reviews.

[20]  P. Nurden,et al.  Platelets and wound healing. , 2008, Frontiers in bioscience : a journal and virtual library.

[21]  F. Marini,et al.  Clarification of the nomenclature for MSC: The International Society for Cellular Therapy position statement. , 2005, Cytotherapy.

[22]  Naoki Ishiguro,et al.  Transplantation of culture expanded bone marrow cells and platelet rich plasma in distraction osteogenesis of the long bones. , 2007, Bone.

[23]  Huipin Yuan,et al.  3D microenvironment as essential element for osteoinduction by biomaterials. , 2005, Biomaterials.

[24]  E. Luong-van,et al.  The in vivo assessment of a novel scaffold containing heparan sulfate for tissue engineering with human mesenchymal stem cells , 2007, Journal of Molecular Histology.

[25]  A. Ratcliffe,et al.  Growth factor-rich plasma increases tendon cell proliferation and matrix synthesis on a synthetic scaffold: an in vitro study. , 2010, Tissue engineering. Part A.

[26]  L Geris,et al.  Current views on calcium phosphate osteogenicity and the translation into effective bone regeneration strategies. , 2012, Acta biomaterialia.

[27]  M. Pittenger,et al.  Multilineage potential of adult human mesenchymal stem cells. , 1999, Science.

[28]  D. Bates,et al.  The Role of Vascular Endothelial Growth Factor in Wound Healing , 2003, The international journal of lower extremity wounds.

[29]  J. Schrooten,et al.  Mechanisms of ectopic bone formation by human osteoprogenitor cells on CaP biomaterial carriers. , 2012, Biomaterials.

[30]  E. Jones,et al.  The role of mesenchymal stem cells in maintenance and repair of bone. , 2007, Injury.

[31]  Antal Rot,et al.  Platelet-released supernatants increase migration and proliferation, and decrease osteogenic differentiation of bone marrow-derived mesenchymal progenitor cells under in vitro conditions , 2004, Platelets.

[32]  Blagoy Blagoev,et al.  Mechanism of Divergent Growth Factor Effects in Mesenchymal Stem Cell Differentiation , 2005, Science.

[33]  M. Jansson,et al.  Fetal Mesenchymal Stem-Cell Engraftment in Bone after In Utero Transplantation in a Patient with Severe Osteogenesis Imperfecta , 2005, Transplantation.

[34]  T. Lapidot,et al.  Stromal-derived factor-1 promotes the growth, survival, and development of human bone marrow stromal stem cells. , 2005, Blood.

[35]  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.

[36]  S. Llames,et al.  Human plasma as a dermal scaffold for the generation of a completely autologous bioengineered skin , 2004, Transplantation.

[37]  Paolo Giannoni,et al.  A tissue engineering approach to bone repair in large animal models and in clinical practice. , 2007, Biomaterials.

[38]  Robert E Guldberg,et al.  Combination of platelet-rich plasma with polycaprolactone-tricalcium phosphate scaffolds for segmental bone defect repair. , 2007, Journal of biomedical materials research. Part A.

[39]  R. Cancedda,et al.  The recruitment of two consecutive and different waves of host stem/progenitor cells during the development of tissue-engineered bone in a murine model. , 2010, Biomaterials.

[40]  Philippe Bierling,et al.  Osteoblastic differentiation of human mesenchymal stem cells with platelet lysate. , 2010, Biomaterials.

[41]  P. Kasten,et al.  Influence of Platelet-Rich Plasma on Osteogenic Differentiation of Mesenchymal Stem Cells and Ectopic Bone Formation in Calcium Phosphate Ceramics , 2006, Cells Tissues Organs.

[42]  Antonios G Mikos,et al.  Pre-culture period of mesenchymal stem cells in osteogenic media influences their in vivo bone forming potential. , 2007, Journal of biomedical materials research. Part A.

[43]  V. Bousson,et al.  Long‐bone critical‐size defects treated with tissue‐engineered grafts: A study on sheep , 2007, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[44]  D. Strunk,et al.  Human Alternatives to Fetal Bovine Serum for the Expansion of Mesenchymal Stromal Cells from Bone Marrow , 2009, Stem cells.

[45]  G. Orive,et al.  Fibroblastic response to treatment with different preparations rich in growth factors , 2009, Cell proliferation.

[46]  H. Sørensen,et al.  Autologous fibrin glue with growth factors in reconstructive maxillofacial surgery. , 2004, International journal of oral and maxillofacial surgery.

[47]  Hans Hauner,et al.  Cartilage-like gene expression in differentiated human stem cell spheroids: a comparison of bone marrow-derived and adipose tissue-derived stromal cells. , 2003, Arthritis and rheumatism.

[48]  J. Lou,et al.  Bone regeneration by implantation of adipose-derived stromal cells expressing BMP-2. , 2007, Biochemical and biophysical research communications.

[49]  R. Cancedda,et al.  The role of bFGF on the ability of MSC to activate endogenous regenerative mechanisms in an ectopic bone formation model. , 2012, Biomaterials.

[50]  J. Zielak,et al.  Platelet-rich plasma diminishes calvarial bone repair associated with alterations in collagen matrix composition and elevated CD34+ cell prevalence. , 2010, Bone.

[51]  K. Kang,et al.  Paracrine effect of canine allogenic umbilical cord blood-derived mesenchymal stromal cells mixed with beta-tricalcium phosphate on bone regeneration in ectopic implantations. , 2010, Cytotherapy.

[52]  E. Vogler,et al.  Contact activation of blood-plasma coagulation. , 2009, Biomaterials.

[53]  W. Mutschler,et al.  Tissue engineering for bone defect healing: an update on a multi-component approach. , 2008, Injury.

[54]  B. Boyan,et al.  Platelet-rich plasma inhibits demineralized bone matrix-induced bone formation in nude mice. , 2007, The Journal of bone and joint surgery. American volume.

[55]  Benjamin M. Wu,et al.  Human adipose-derived stromal cells stimulate autogenous skeletal repair via paracrine Hedgehog signaling with calvarial osteoblasts. , 2011, Stem cells and development.

[56]  X. Holy,et al.  Platelet lysates promote mesenchymal stem cell expansion: A safety substitute for animal serum in cell‐based therapy applications , 2005, Journal of cellular physiology.

[57]  Hideki Yoshikawa,et al.  Circulating Bone Marrow‐Derived Osteoblast Progenitor Cells Are Recruited to the Bone‐Forming Site by the CXCR4/Stromal Cell‐Derived Factor‐1 Pathway , 2008, Stem cells.

[58]  G. Intini.  The use of platelet-rich plasma in bone reconstruction therapy. , 2009, Biomaterials.

[59]  Darwin J. Prockop,et al.  Transplantability and therapeutic effects of bone marrow-derived mesenchymal cells in children with osteogenesis imperfecta , 1999, Nature Medicine.

[60]  A. Dohan,et al.  Platelet-rich fibrin (PRF): a second-generation platelet concentrate. Part I: technological concepts and evolution. , 2006, Oral surgery, oral medicine, oral pathology, oral radiology, and endodontics.

[61]  A. Caplan,et al.  In vitro dexamethasone pretreatment enhances bone formation of human mesenchymal stem cells in vivo , 2009, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[62]  F. Speleman,et al.  Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes , 2002, Genome Biology.

[63]  R. Cancedda,et al.  The development of tissue-engineered bone of different origin through endochondral and intramembranous ossification following the implantation of mesenchymal stem cells and osteoblasts in a murine model. , 2010, Biomaterials.

[64]  C. Choong,et al.  PDGF, TGF-beta, and FGF signaling is important for differentiation and growth of mesenchymal stem cells (MSCs): transcriptional profiling can identify markers and signaling pathways important in differentiation of MSCs into adipogenic, chondrogenic, and osteogenic lineages. , 2008, Blood.

[65]  E H Schemitsch,et al.  Fracture vascularity and bone healing: a systematic review of the role of VEGF. , 2008, Injury.

[66]  E. Anitua.  The use of plasma-rich growth factors (PRGF) in oral surgery. , 2001, Practical procedures & aesthetic dentistry : PPAD.

[67]  R Cancedda,et al.  Repair of large bone defects with the use of autologous bone marrow stromal cells. , 2001, The New England journal of medicine.

[68]  C. Marchetti,et al.  Mesenchymal stem cells and platelet-rich plasma enhance bone formation in sinus grafting: a histomorphometric study in minipigs. , 2008, Journal of clinical periodontology.

[69]  M. Betsch,et al.  Combined use of platelet-rich plasma and autologous bone grafts in the treatment of long bone defects in mini-pigs. , 2010, Injury.

[70]  G. Evans,et al.  In vivo osteogenic potential of human adipose-derived stem cells/poly lactide-co-glycolic acid constructs for bone regeneration in a rat critical-sized calvarial defect model. , 2007, Tissue engineering.