Guiding Chondrogenesis and Osteogenesis with Mineral-Coated Hydroxyapatite and BMP-2 Incorporated within High-Density hMSC Aggregates for Bone Regeneration.

Since hydroxyapatite and bone morphogenetic protein-2 (BMP-2) can regulate chondrogenesis and osteogenesis, their individual and combined effects on endochondral ossification within human bone marrow-derived stem cell (hMSC) aggregates were investigated. Hydroxyapatite was presented in the form of mineral-coated hydroxyapatite microparticles (MCM) capable of controlled BMP-2 delivery. Aggregates were treated with varied BMP-2 concentrations supplemented in the media and loaded onto MCM to examine the influence of BMP-2 amount and spatial presentation on regulating chondrogenesis and osteogenesis. MCM alone induced GAG and type II collagen production by week 5 for two of three donors, and BMP-2 may have accelerated MCM-induced chondrogenesis. ALP activity and calcium content of cells-only aggregates suggest that the BMP-2-induced osteogenic response may be concentration-dependent. Treatment with MCM and BMP-2 resulted in chondrogenesis as early as week 2, which may have promoted additional mineralization by week 5, suggesting the induction of endochondral ossification. Released BMP-2 had similar if not higher levels of bioactivity compared to that of exogenous BMP-2 with regard to chondrogenesis and osteogenesis. In addition to providing localized and sustained BMP-2 delivery, MCM incorporation within aggregates yields a self-sustaining system that may be injected or implanted more rapidly to heal bone defects through endochondral ossification without extended in vitro culture.

[1]  Eben Alsberg,et al.  Multilayered Inorganic Microparticles for Tunable Dual Growth Factor Delivery , 2014, Advanced functional materials.

[2]  H. Weinans,et al.  Chondrogenically differentiated mesenchymal stromal cell pellets stimulate endochondral bone regeneration in critical-sized bone defects. , 2014, European cells & materials.

[3]  Xianting Ding,et al.  Guiding the osteogenic fate of mouse and human mesenchymal stem cells through feedback system control , 2013, Scientific Reports.

[4]  Jerry C. Hu,et al.  Self-organization and the self-assembling process in tissue engineering. , 2013, Annual review of biomedical engineering.

[5]  E. Alsberg,et al.  High-density cell systems incorporating polymer microspheres as microenvironmental regulators in engineered cartilage tissues. , 2013, Tissue engineering. Part B, Reviews.

[6]  M. Reinders,et al.  Predicting the therapeutic efficacy of MSC in bone tissue engineering using the molecular marker CADM1. , 2013, Biomaterials.

[7]  A. Jahanshahi,et al.  Study on nano-structured hydroxyapatite/zirconia stabilized yttria on healing of articular cartilage defect in rabbit. , 2013, Acta cirurgica brasileira.

[8]  Siew Lok Toh,et al.  Enhanced osteoinductivity and osteoconductivity through hydroxyapatite coating of silk-based tissue-engineered ligament scaffold. , 2013, Journal of biomedical materials research. Part A.

[9]  Kevin P Egan,et al.  Bone histomorphometry using free and commonly available software , 2012, Histopathology.

[10]  E. Alsberg,et al.  Spatiotemporal Regulation of Chondrogenic Differentiation with Controlled Delivery of Transforming Growth Factor‐β1 from Gelatin Microspheres in Mesenchymal Stem Cell Aggregates , 2012, Stem cells translational medicine.

[11]  Rocky S Tuan,et al.  Concise Review: The Clinical Application of Mesenchymal Stem Cells for Musculoskeletal Regeneration: Current Status and Perspectives , 2012, Stem cells translational medicine.

[12]  H. Thielecke,et al.  A scaffold-free in vitro model for osteogenesis of human mesenchymal stem cells. , 2011, Tissue & cell.

[13]  F. O'Brien,et al.  In-vivo generation of bone via endochondral ossification by in-vitro chondrogenic priming of adult human and rat mesenchymal stem cells , 2011, BMC musculoskeletal disorders.

[14]  C. Beltrami,et al.  Biochemical and biophysical analyses of tissue-engineered bone obtained from three-dimensional culture of a subset of bone marrow mesenchymal stem cells. , 2010, Tissue engineering. Part A.

[15]  Farshid Guilak,et al.  Isolation of adipose-derived stem cells and their induction to a chondrogenic phenotype , 2010, Nature Protocols.

[16]  M. Kassem,et al.  Parameters in three-dimensional osteospheroids of telomerized human mesenchymal (stromal) stem cells grown on osteoconductive scaffolds that predict in vivo bone-forming potential. , 2010, Tissue engineering. Part A.

[17]  Ivan Martin,et al.  Recapitulation of endochondral bone formation using human adult mesenchymal stem cells as a paradigm for developmental engineering , 2010, Proceedings of the National Academy of Sciences.

[18]  B. Wildemann,et al.  Bone morphogenetic proteins in critical-size bone defects: what are the options? , 2009, Injury.

[19]  W. Murphy,et al.  Growth of hydroxyapatite coatings on biodegradable polymer microspheres. , 2009, ACS applied materials & interfaces.

[20]  C. H. Coyle,et al.  Sustained hypoxia enhances chondrocyte matrix synthesis , 2009, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[21]  Hwan-You Chang,et al.  Recent advances in three‐dimensional multicellular spheroid culture for biomedical research , 2008, Biotechnology journal.

[22]  Yilin Cao,et al.  The dependence of in vivo stable ectopic chondrogenesis by human mesenchymal stem cells on chondrogenic differentiation in vitro. , 2008, Biomaterials.

[23]  D. P. O'connor,et al.  Outcomes of Tibial Nonunion in Older Adults Following Treatment Using the Ilizarov Method , 2007, Journal of orthopaedic trauma.

[24]  C. V. van Blitterswijk,et al.  Donor variation and loss of multipotency during in vitro expansion of human mesenchymal stem cells for bone tissue engineering , 2007, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[25]  J. Welter,et al.  Simplification of aggregate culture of human mesenchymal stem cells as a chondrogenic screening assay. , 2007, BioTechniques.

[26]  T. Turner,et al.  Additive enhancement of implant fixation following combined treatment with rhTGF-beta2 and rhBMP-2 in a canine model. , 2006, The Journal of bone and joint surgery. American volume.

[27]  M. Baumann,et al.  Hepatocyte growth factor (HGF) adsorption kinetics and enhancement of osteoblast differentiation on hydroxyapatite surfaces. , 2005, Biomaterials.

[28]  M. Manley,et al.  Hydroxyapatite-coated prostheses in total hip and knee arthroplasty. , 2004, The Journal of bone and joint surgery. American volume.

[29]  V. Alt,et al.  Effect of glycerol-L-lactide coating polymer on bone ingrowth of bFGF-coated hydroxyapatite implants. , 2004, Journal of controlled release : official journal of the Controlled Release Society.

[30]  C. Kaps,et al.  BMP2 initiates chondrogenic lineage development of adult human mesenchymal stem cells in high-density culture. , 2003, Differentiation; research in biological diversity.

[31]  M. Mastrogiacomo,et al.  Formation of a chondro-osseous rudiment in micromass cultures of human bone-marrow stromal cells , 2003, Journal of Cell Science.

[32]  Richard Tuli,et al.  Adult mesenchymal stem cells and cell-based tissue engineering , 2002, Arthritis research & therapy.

[33]  S. Santavirta,et al.  Recombinant Human Bone Morphogenetic Protein-2 for Treatment of Open Tibial Fractures: A Prospective, Controlled, Randomized Study of Four Hundred and Fifty Patients , 2002, The Journal of bone and joint surgery. American volume.

[34]  T. Pufe,et al.  Expression of VEGF121 and VEGF165 in hypertrophic chondrocytes of the human growth plate and epiphyseal cartilage , 2002, Journal of anatomy.

[35]  A. Scutt,et al.  Histochemical Localization of Alkaline Phosphatase Activity in Decalcified Bone and Cartilage , 2002, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[36]  D J Prockop,et al.  Donor variation in the growth properties and osteogenic potential of human marrow stromal cells , 1999, Journal of cellular biochemistry.

[37]  A I Caplan,et al.  In vitro chondrogenesis of bone marrow-derived mesenchymal progenitor cells. , 1998, Experimental cell research.

[38]  T. Sullivan,et al.  Rapid chondrocyte maturation by serum‐free culture with BMP‐2 and ascorbic acid , 1997, Journal of cellular biochemistry.

[39]  M J Yaszemski,et al.  Bone formation by three-dimensional stromal osteoblast culture in biodegradable polymer scaffolds. , 1997, Journal of biomedical materials research.

[40]  M J Yaszemski,et al.  Ectopic bone formation by marrow stromal osteoblast transplantation using poly(DL-lactic-co-glycolic acid) foams implanted into the rat mesentery. , 1997, Journal of biomedical materials research.

[41]  I. Kangasniemi,et al.  Subchondral bone and cartilage repair with bioactive glasses, hydroxyapatite, and hydroxyapatite-glass composite. , 1996, Journal of biomedical materials research.

[42]  M. Habal,et al.  Bone grafts and bone induction substitutes. , 1994, Clinics in plastic surgery.

[43]  V. Rosen,et al.  Recombinant human bone morphogenetic protein induces bone formation. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[44]  V. Rosen,et al.  Novel regulators of bone formation: molecular clones and activities. , 1988, Science.

[45]  A. Grodzinsky,et al.  Fluorometric assay of DNA in cartilage explants using Hoechst 33258. , 1988, Analytical biochemistry.

[46]  D. Buttle,et al.  Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. , 1986, Biochimica et biophysica acta.

[47]  E. White,et al.  The restoration of the articular surfaces overlying Replamineform porous biomaterials. , 1977, Journal of biomedical materials research.

[48]  G. Canton,et al.  Clinical applications of growth factors in bone injuries: experience with BMPs. , 2013, Injury.

[49]  N. Kübler,et al.  Scaffold-free microtissues: differences from monolayer cultures and their potential in bone tissue engineering , 2012, Clinical Oral Investigations.

[50]  Cartilage Biology.  Hypertrophy in Mesenchymal Stem Cell Chondrogenesis: Effect of TGF- Isoforms and Chondrogenic Conditioning , 2010 .

[51]  E. Schipani.  Hypoxia and HIF-1alpha in chondrogenesis. , 2006, Annals of the New York Academy of Sciences.

[52]  P. Giannoudis,et al.  Impact of hypothermia on the immunologic response after trauma and elective surgery. , 2005, Surgical technology international.

[53]  C. Bünger,et al.  A review of ceramic coatings for implant fixation. , 1999, Journal of long-term effects of medical implants.

[54]  A I Caplan,et al.  Characterization of cells with osteogenic potential from human marrow. , 1992, Bone.