Integration of a novel injectable nano calcium sulfate/alginate scaffold and BMP2 gene-modified mesenchymal stem cells for bone regeneration

Tissue Engineering - Part A

Volumn 19, Issue 3-4, 2013, Pages 508-518

  He, Xiaoning ^a,b   Dziak, Rosemary ^a   Mao, Keya ^c   Genco, Robert ^a   Swithart, Mark ^d   Li, Chunyi ^a   Yang, Shuying ^a  

^a UNIVERSITY AT BUFFALO   (United States)

^b CHINA MEDICAL UNIVERSITY   (China)

^c CHINESE PLA GENERAL HOSPITAL   (China)

^d the State University of New York   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ABILITY TESTING; BIOCOMPATIBILITY; BLOOD VESSELS; BONE; DEFECTS; FLOWCHARTING; GENES; MECHANICAL PROPERTIES; PHOSPHATASES; REPAIR; SCAFFOLDS (BIOLOGY);

ALKALINE PHOSPHATASE ACTIVITY; ANATOMICAL STRUCTURES; BLOOD VESSEL GROWTH; BONE DEFECT; BONE FORMATION; BONE MORPHOGENETIC PROTEIN-2; BONE REGENERATION; BONE TISSUE ENGINEERING; CALCIUM SULFATE; CALVARIAL BONES; CRANIOFACIAL; CRANIOFACIAL SKELETON; INJECTION FORCE; MESENCHYMAL STEM CELL; MINIMALLY INVASIVE SURGERY; NANO SCALE; OSTEOCONDUCTIVE; OSTEOGENIC ACTIVITY; RAT MODEL; VASCULARIZATION;

CELL CULTURE;

ANIMALIA; RATTUS;

ALGINIC ACID; BMP2 PROTEIN, RAT; BONE MORPHOGENETIC PROTEIN 2; CALCIUM SULFATE; NANOCAPSULE; TISSUE SCAFFOLD;

ANIMAL; ARTICLE; BONE REGENERATION; CHEMISTRY; DRUG EFFECT; DRUG IMPLANT; EQUIPMENT; EQUIPMENT DESIGN; EQUIPMENT FAILURE; INJECTION; MALE; MESENCHYMAL STEM CELL TRANSPLANTATION; MULTIMODALITY CANCER THERAPY; PHYSIOLOGY; RAT; SKULL FRACTURE; SPRAGUE DAWLEY RAT; TREATMENT OUTCOME;

ALGINATES; ANIMALS; BONE MORPHOGENETIC PROTEIN 2; BONE REGENERATION; CALCIUM SULFATE; COMBINED MODALITY THERAPY; DRUG IMPLANTS; EQUIPMENT DESIGN; EQUIPMENT FAILURE ANALYSIS; INJECTIONS; MALE; MESENCHYMAL STEM CELL TRANSPLANTATION; NANOCAPSULES; RATS; RATS, SPRAGUE-DAWLEY; SKULL FRACTURES; TISSUE SCAFFOLDS; TREATMENT OUTCOME;

MLCS; MLOWN;

EID: 84871080241     PISSN: 19373341     EISSN: 1937335X     Source Type: Journal    
DOI: 10.1089/ten.tea.2012.0244     Document Type: Article

References (99)

1. Bowers, G.M., Chadroff, B., Carnevale, R., Mellonig, J., Corio, R., Emerson, J., et al. Histologic evaluation of new attachment apparatus formation in humans. Part III. J Peri-odontol 60, 683, 1989.
2. Laurell, L., Gottlow, J., Zybutz, M., and Persson, R. Treat-ment of intrabony defects by different surgical procedures. A literature review. J Periodontol 69, 303, 1998.
3. Samstein, B., and Platt, J.L. Physiologic and immunologic hurdles to xenotransplantation. J Am Soc Nephrol 12, 182, 2001.
4. Zuk, P.A. Tissue engineering craniofacial defects with adult stem cells? Are we ready yet? Pediatr Res 63, 478, 2008.
5. Li, Z., Ramay, H.R., Hauch, K.D., Xiao, D., and Zhang, M. Chitosan-alginate hybrid scaffolds for bone tissue engi-neering. Biomaterials 26, 3919, 2005.
6. Laurencin, C.T., Ambrosio, A.M., Borden, M.D., and Coop-er, J.A., Jr. Tissue engineering: orthopedic applications. Annu Rev Biomed Eng 1, 19, 1999.
7. Kretlow, J.D., Young, S., Klouda, L., Wong, M., and Mikos, A.G. Injectable biomaterials for regenerating complex cra-niofacial tissues. Adv Mater 21, 3368, 2009.
8. Xu, H.H., Weir, M.D., and Simon, C.G. Injectable and strong nano-apatite scaffolds for cell/growth factor delivery and bone regeneration. Dent Mater 24, 1212, 2008.
9. Drury, J.L., and Mooney, D.J. Hydrogels for tissue engi-neering: scaffold design variables and applications. Bioma-terials 24, 4337, 2003.
10. Peters, C.L., Hines, J.L., Bachus, K.N., Craig, M.A., and Bloebaum, R.D. Biological effects of calcium sulfate as a bone graft substitute in ovine metaphyseal defects. J Biomed Mater Res Part A 76, 456, 2006.
11. Maeda, S.T., Bramane, C.M., Taga, R., Garcia, R.B., de Moraes, I.G., and Bernadineli, N. Evaluation of surgical cavities filled with three types of calcium sulfate. J Appl Oral Sci 15, 416, 2007.
12. Kim, J.H., Oh, J.H., Han, I., Kim, H.S., and Chung, S.W. Grafting using injectable calcium sulfate in bone tumor surgery: comparison with demineralized bone matrix-based grafting. Clin Orthop Surg 3, 191, 2011.
13. Qi, Y., Wang, Y., Yan, W., Li, H., Shi, Z., and Pan, Z. Combined mesenchymal stem cell sheets and rhBMP-2-releasing calcium-sulfate-rhBMP-2 scaffolds for segmental bone tissue engineering. Cell Transplant 21, 693, 2012.
14. Zhao, W., Chang, J., and Zhai, W. Self-setting properties and in vitro bioactivity of Ca3SiO5/CaSO4·1/2H2O composite cement. J Biomed Mater Res A 85, 336, 2008.
15. Chen, Z., Liu, H., Liu, X., and Cui, F.Z. Injectable calcium sulfate/mineralized collagen-based bone repair materials with regulable self-setting properties. J Biomed Mater Res A 99, 554, 2011.
16. Cho, B.C., Park, J.W., Baik, B.S., and Kim, I.S. Clinical ap-plication of injectable calcium sulfate on early bony consol-idation in distraction osteogenesis for the treatment of craniofacial microsomia. J Craniofac Surg 13, 465; discussion 75, 2002.
17. Murashima, Y., Yoshikawa, G., Wadachi, R., Sawada, N., and Suda, H. Calcium sulphate as a bone substitute for various osseous defects in conjunction with apicectomy. Int Endod J 35, 768, 2002.
18. Sukumar, S., Drizhal, I., Paulusova, V., and Bukac, J. Surgical treatment of periodontal intrabony defects with calcium sulphate in combination with beta-tricalcium phosphate: clinical observations two years post-surgery. Acta Medica (Hradec Kralove) 54, 13, 2011.
19. Pecora, G., Andreana, S., Margarone, J.E., 3rd, Covani, U., and Sottosanti, J.S. Bone regeneration with a calcium sulfate barrier. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 84, 424, 1997.
20. Zhang, S. Fabrication of novel biomaterials through molec-ular self-assembly. Nat Biotechnol 21, 1171, 2003.
21. Stupp, S.I., and Braun, P.V. Molecular manipulation of mi-crostructures: biomaterials, ceramics, and semiconductors. Science 277, 1242, 1997.
22. Kong, L.X., Peng, Z., Li, S.D., and Bartold, P.M. Nano-technology and its role in the management of periodontal diseases. Periodontol 2000 40, 184, 2006.
23. Park, Y.B., Mohan, K., Al-Sanousi, A., Almaghrabi, B., Genco, R.J., Swihart, M.T., et al. Synthesis and characteriza-tion of nanocrystalline calcium sulfate for use in osseous regeneration. Biomed Mater 6, 055007, 2011.
24. Mastrogiacomo, M., Muraglia, A., Komlev, V., Peyrin, F., Rustichelli, F., Crovace, A., et al. Tissue engineering of bone: search for a better scaffold. Orthod Craniofac Res 8, 277, 2005.
25. Hollister, S.J. Porous scaffold design for tissue engineering. Nat Mater 4, 518, 2005.
26. Lin, H.R., and Yeh, Y.J. Porous alginate/hydroxyapatite composite scaffolds for bone tissue engineering: preparation, characterization, and in vitro studies. J Biomed Mater Res B Appl Biomater 71, 52, 2004.
27. Grandi, C., Di Liddo, R., Paganin, P., Lora, S., Dalzoppo, D., Feltrin, G., et al. Porous alginate/poly(epsilon-caprolactone) scaffolds: preparation, characterization and in vitro biologi-cal activity. Int J Mol Med 27, 455, 2011.
28. Shang, Q., Wang, Z., Liu, W., Shi, Y., Cui, L., and Cao, Y. Tissue-engineered bone repair of sheep cranial defects with autologous bone marrow stromal cells. J Craniofac Surg 12, 586; discussion 94, 2001.
29. Chang, S.C., Rowley, J.A., Tobias, G., Genes, N.G, Roy, A.K., Mooney, D.J., et al. Injection molding of chondrocyte/ alginate constructs in the shape of facial implants. J Biomed Mater Res 55, 503, 2001.
30. Thomas, S. Alginate dressings in surgery and wound man-agement\-Part 1. J Wound Care 9, 56, 2000.
31. Smidsrod, O., and Skjak-Braek, G. Alginate as immobiliza-tion matrix for cells. Trends Biotechnol 8, 71, 1990.
32. Matthew, I.R., Browne, R.M., Frame, J.W., and Millar, B.G Subperiosteal behaviour of alginate and cellulose wound dressing materials. Biomaterials 16, 275, 1995.
33. Wang, L., Shelton, R.M., Cooper, P.R., Lawson, M., Triffitt, J.T., and Barralet, J.E. Evaluation of sodium alginate for bone marrow cell tissue engineering. Biomaterials 24, 3475, 2003.
34. Dobratz, E.J., Kim, S.W., Voglewede, A., and Park, S.S. In-jectable cartilage: using alginate and human chondrocytes. Arch Facial Plast Surg 11, 40, 2009.
35. Oest, M.E., Dupont, K.M., Kong, H.J., Mooney, D.J., and Guldberg, R.E. Quantitative assessment of scaffold and growth factor-mediated repair of critically sized bone defects. J Orthop Res 25, 941, 2007.
36. Grant, G.T., Morris, E.R., Rees, D.A., Smith, P.J.C., and Thom, D. Biological interactions between polysaccharides and divalent cations - egg-box model. FEBS Lett 32, 195, 1973.
37. Stokke, B.T., Smidsrod, O., Bruheim, P., and Skjakbraek, G. Distribution of uronate residues in alginate chains in relation to alginate gelling properties. Macromolecules 24, 4637, 1991.
38. Lee, G.S., Park, J.H., Shin, U.S., and Kim, H.W. Direct de-posited porous scaffolds of calcium phosphate cement with alginate for drug delivery and bone tissue engineering. Acta Biomater 7, 3178, 2011.
39. Wee, S., and Gombotz, W.R. Protein release from alginate matrices. Adv Drug Deliv Rev 31, 267, 1998.
40. Tan, R., Feng, Q., Jin, H., Li, J., Yu, X., She, Z., et al. Structure and biocompatibility of an injectable bone regeneration composite. J Biomater Sci Polym Ed 22, 1861, 2011.
41. Hyland, L.L., Taraban, M.B., Hammouda, B., and Bruce Yu, Y. Mutually reinforced multicomponent polysaccharide networks. Biopolymers 95, 840, 2011.
42. Bruder, S.P., Kraus, K.H., Goldberg, V.M., and Kadiyala, S. The effect of implants loaded with autologous mesenchymal stem cells on the healing of canine segmental bone defects. J Bone Joint Surg Am 80, 985, 1998.
43. Pittenger, M.F., Mackay, A.M., Beck, S.C., Jaiswal, R.K., Douglas, R., Mosca, J.D., et al. Multilineage potential of adult human mesenchymal stem cells. Science 284, 143, 1999.
44. Muschler, G.F., Nakamoto, C., and Griffith, L.G. Engineer-ing principles of clinical cell-based tissue engineering. J Bone Joint Surg Am 86-A, 1541, 2004.
45. Wang, C., Wang, Z., Li, A., Bai, F., Lu, J., Xu, S., et al. Repair of segmental bone-defect of goat's tibia using a dynamic perfusion culture tissue engineering bone. J Biomed Mater Res A 92, 1145, 2010.
46. Azad, V., Breitbart, E., Al-Zube, L., Yeh, S., O'Connor, J.P., and Lin, S.S. rhBMP-2 enhances the bone healing response in a diabetic rat segmental defect model. J Orthop Trauma 23, 267, 2009.
47. Jones, A.L., Bucholz, R.W., Bosse, M.J., Mirza, S.K., Lyon, T.R., Webb, L.X., et al. Recombinant human BMP-2 and allograft compared with autogenous bone graft for reconstruction of diaphyseal tibial fractures with cortical defects. A randomized, controlled trial. J Bone Joint Surg Am 88, 1431, 2006.
48. Gautschi, O.P., Frey, S.P., and Zellweger, R Bone morphoge-netic proteins in clinical applications. ANZ J Surg 77,626,2007.
49. Yue, B., Lu, B., Dai, K.R., Zhang, X.L., Yu, C.F., Lou, J.R., et al. BMP2 gene therapy on the repair of bone defects of aged rats. Calcif Tissue Int 77, 395, 2005.
50. Park, Y., Dziak, R., Genco, R., Swihart, M.T., and Periopa-nayagam, H. Calcium Sulfate Based Nanoparticles. US Patent: US7767226.
51. Salvadori, B., Capitani, G., Mellini, M., and Dei, L. A novel method to prepare inorganic water-soluble nanocrystals. J Colloid Interface Sci 298, 487, 2006.
52. Khairoun, I., Boltong, M.G., Driessens, F.C., and Planell, JA. Some factors controlling the injectability of calcium phos-phate bone cements. J Mater Sci Mater Med 9, 425, 1998.
53. Ginebra, M.P., Rilliard, A., Fernandez, E., Elvira, C., San Roman, J., and Planell, J.A. Mechanical and rheological im-provement of a calcium phosphate cement by the addition of a polymeric drug. J Biomed Mater Res 57, 113, 2001.
54. Xu, H.H., and Quinn, J.B. Calcium phosphate cement con-taining resorbable fibers for short-term reinforcement and macroporosity. Biomaterials 23, 193, 2002.
55. Xu, H.H., Takagi, S., Quinn, J.B., and Chow, L.C. Fast-setting calcium phosphate scaffolds with tailored macropore for-mation rates for bone regeneration. J Biomed Mater Res Part A 68, 725, 2004.
56. Xu, H.H., Eichmiller, F.C., and Giuseppetti, A.A. Re-inforcement of a self-setting calcium phosphate cement with different fibers. J Biomed Mater Res 52, 107, 2000.
57. Franceschi, R.T., Yang, S., Rutherford, R.B., Krebsbach, P.H., Zhao, M., and Wang, D. Gene therapy approaches for bone regeneration. Cells Tissues Organs 176, 95, 2004.
58. Yang, S., Wei, D., Wang, D., Phimphilai, M., Krebsbach, P.H., and Franceschi, R.T. In vitro and in vivo synergistic interactions between the Runx2/Cbfa1 transcription factor and bone morphogenetic protein-2 in stimulating osteoblast differentiation. J Bone Miner Res 18, 705, 2003.
59. Schneider, G.B., Whitson, S.W., and Cooper, L.F. Restricted and coordinated expression of beta3-integrin and bone sia-loprotein during cultured osteoblast differentiation. Bone 24, 321, 1999.
60. Yang, S., and Li, Y.P. RGS10-null mutation impairs osteo-clast differentiation resulting from the loss of [Ca2+]i oscil-lation regulation. Genes Dev 21, 1803, 2007.
61. Barzilay, R., Sadan, O., Melamed, E., and Offen, D. Comparative characterization of bone marrow-derived mesenchymal stromal cells from four different rat strains. Cytotherapy 11, 435, 2009.
62. Harting, M., Jimenez, F., Pati, S., Baumgartner, J., and Cox, C., Jr. Immunophenotype characterization of rat mesenchy-mal stromal cells. Cytotherapy 10, 243, 2008.
63. McCarty, R.C., Gronthos, S., Zannettino, A.C., Foster, B.K., and Xian, C.J. Characterisation and developmental potential of ovine bone marrow derived mesenchymal stem cells. J Cell Physiol 219, 324, 2009.
64. Ruggeri, Z.M. von Willebrand factor. J Clin Invest 100, S41, 1997.
65. Vailhe, B., Dietl, J., Kapp, M., Toth, B., and Arck, P. In-creased blood vessel density in decidua parietalis is associ-ated with spontaneous human first trimester abortion. Hum Reprod 14, 1628, 1999.
66. Reddy, S., Wasnik, S., Guha, A., Kumar, J.M., Sinha, A., and Singh, S. Evaluation of nano-biphasic calcium phosphate ceramics for bone tissue engineering applications: in vitro and preliminary in vivo studies. J Biomater Appl 2012. DOI: 10.1177/0885328211415132
67. Moreau, J.L., and Xu, H.H. Mesenchymal stem cell pro-liferation and differentiation on an injectable calcium phosphate-chitosan composite scaffold. Biomaterials 30, 2675, 2009.
68. Nan, K., Sun, S., Li, Y., Chen, H., Wu, T., and Lu, F. Ectopic osteogenic ability of calcium phosphate scaffolds cultured with osteoblasts. J Biomed Mater Res A 93, 464, 2010.
69. Yuan, H., van Blitterswijk, C.A., de Groot, K., and de Bruijn, J.D. Cross-species comparison of ectopic bone formation in biphasic calcium phosphate (BCP) and hydroxyapatite (HA) scaffolds. Tissue Eng 12, 1607, 2006.
70. Hollinger, J.O., and Battistone, G.C. Biodegradable bone repair materials. Synthetic polymers and ceramics. Clin Orthop Relat Res 290, 1986.
71. Garg, T., Singh, O., Arora, S., and Murthy, R Scaffold: a novel carrier for cell and drug delivery. Crit Rev Ther Drug Carrier Syst 29, 1, 2012.
72. Yamada, Y., Ueda, M., Naiki, T., Takahashi, M., Hata, K., and Nagasaka, T. Autogenous injectable bone for regenera-tion with mesenchymal stem cells and platelet-rich plasma: tissue-engineered bone regeneration. Tissue Eng 10, 955, 2004.
73. Orsini, M., Orsini, G., Benlloch, D., Aranda, J.J., Lazaro, P., Sanz, M., et al. Comparison of calcium sulfate and autoge-nous bone graft to bioabsorbable membranes plus autoge-nous bone graft in the treatment of intrabony periodontal defects: a split-mouth study. J Periodontol 72, 296, 2001.
74. Bron, J.L., Vonk, L.A., Smit, T.H., and Koenderink, G.H. Engineering alginate for intervertebral disc repair. J Mech Behav Biomed Mater 4, 1196, 2011.
75. Park, D.J., Choi, B.H., Zhu, S.J., Huh, J.Y., Kim, B.Y., and Lee, S.H. Injectable bone using chitosan-alginate gel/ mesenchymal stem cells/BMP-2 composites. J Craniomax-illofac Surg 33, 50, 2005.
76. Zhao, L., Weir, M.D., and Xu, H.H. An injectable calcium phosphate-alginate hydrogel-umbilical cord mesenchymal stem cell paste for bone tissue engineering. Biomaterials 31, 6502, 2010.
77. Bohari, S.P., Hukins, D.W., and Grover, L.M. Effect of calcium alginate concentration on viability and proliferation of encapsulated fibroblasts. Biomed Mater Eng 21,159, 2011.
78. Gaetani, R., Doevendans, P.A., Metz, C.H., Alblas, J., Messina, E., Giacomello, A., et al. Cardiac tissue engineering using tissue printing technology and human cardiac pro-genitor cells. Biomaterials 33, 1782, 2012.
79. Itala, A.I., Ylanen, H.O., Ekholm, C., Karlsson, K.H., and Aro, H.T. Pore diameter of more than 100 microm is not requisite for bone ingrowth in rabbits. J Biomed Mater Res 58, 679, 2001.
80. Schliephake, H., Neukam, F.W., and Klosa, D. Influence of pore dimensions on bone ingrowth into porous hydroxyl-apatite blocks used as bone graft substitutes. A histometric study. Int J Oral Maxillofac Surg 20, 53, 1991.
81. Harris, W.H., and Jasty, M. Bone ingrowth into porous coated canine acetabular replacements: the effect of pore size, apposition, and dislocation. Hip 214, 1985.
82. Martens, M., Ducheyne, P., De Meester, P., and Mulier, J.C. Skeletal fixation of implants by bone ingrowth into surface pores. Arch Orthop Trauma Surg 97, 111, 1980.
83. Hulbert, S.F., Young, F.A., Mathews, R.S., Klawitter, J.J., Talbert, C.D., and Stelling, F.H. Potential of ceramic mate-rials as permanently implantable skeletal prostheses. J Biomed Mater Res 4, 433, 1970.
84. Bobyn, J.D., Pilliar, R.M., Cameron, H.U., and Weatherly, G.C. The optimum pore size for the fixation of porous-surfaced metal implants by the ingrowth of bone. Clin Orthop Relat Res 263, 1980.
85. Wang, E.A., Rosen, V., D'Alessandro, J.S., Bauduy, M., Cordes, P., Harada, T., et al. Recombinant human bone morphogenetic protein induces bone formation. Proc Natl Acad Sci U S A 87, 2220, 1990.
86. Sandhu, H.S. Bone morphogenetic proteins and spinal sur-gery. Spine (Phila Pa 1976) 28, S64, 2003.
87. Khan, S.N., and Lane, J.M. The use of recombinant human bone morphogenetic protein-2 (rhBMP-2) in orthopaedic applications. Expert Opin Biol Ther 4, 741, 2004.
88. Vaidya, R., Carp, J., Sethi, A., Bartol, S., Craig, J., and Les, C.M. Complications of anterior cervical discectomy and fusion using recombinant human bone morphogenetic protein-2. Eur Spine J 16, 1257, 2007.
89. Marson, A.G., Hutton, J.L., Leach, J.P., Castillo, S., Schmidt, D., White, S., et al. Levetiracetam, oxcarbazepine, remacemide and zonisamide for drug resistant localization-related epi-lepsy: a systematic review. Epilepsy Res 46, 259, 2001.
90. Steinert, A.F., Palmer, G.D., Pilapil, C., Noth, U., Evans, C.H., and Ghivizzani, S.C. Enhanced in vitro chondrogenesis of primary mesenchymal stem cells by combined gene transfer. Tissue Eng Part A 15, 1127, 2009.
91. Castro-Govea, Y., Cervantes-Kardasch, V.H., Borrego-Soto, G., Martinez-Rodriguez, H.G., Espinoza-Juarez, M., Romero-Diaz, V., et al. Human bone morphogenetic pro-tein 2-transduced mesenchymal stem cells improve bone regeneration in a model of mandible distraction surgery. J Craniofac Surg 23, 392, 2012.
92. Sweeney, T.M., Opperman, L.A., Persing, J.A., and Ogle, R.C. Repair of critical size rat calvarial defects using extra-cellular matrix protein gels. J Neurosurg 83, 710, 1995.
93. Rai, B., Oest, M.E., Dupont, K.M., Ho, K.H., Teoh, S.H., and Guldberg, R.E. Combination of platelet-rich plasma with polycaprolactone-tricalcium phosphate scaffolds for seg-mental bone defect repair. J Biomed Mater Res A 81, 888, 2007.
94. Li, Z.H., Liao, W., Cui, X.L., Zhao, Q., Liu, M., Chen, Y.H., et al. Intravenous transplantation of allogeneic bone marrow mesenchymal stem cells and its directional migration to the necrotic femoral head. Int J Med Sci 8, 74, 2011.
95. Saito, T., Kuang, J.Q., Bittira, B., Al-Khaldi, A., and Chiu, R.C. Xenotransplant cardiac chimera: immune tolerance of adult stem cells. Ann Thorac Surg 74, 19; discussion 24, 2002.
96. Devine, S.M., Cobbs, C., Jennings, M., Bartholomew, A., and Hoffman, R. Mesenchymal stem cells distribute to a wide range of tissues following systemic infusion into nonhuman primates. Blood 101, 2999, 2003.
97. Inoue, S., Popp, F.C., Koehl, G.E., Piso, P., Schlitt, H.J., Geissler, E.K., et al. Immunomodulatory effects of mesen-chymal stem cells in a rat organ transplant model. Trans-plantation 81, 1589, 2006.
98. Suwa, F., Fang, Y.R., Toda, I., Tang, C.S., Yang, L.J., Gao, Y.H., et al. SEM study on microvascular changes following implantation of bone morphogenetic protein combined with hydroxyapatite into experimental bone defects. J Osaka Dent Univ 32, 27, 1998.
99. Deckers, M.M., van Bezooijen, R.L., van der Horst, G., Hoogendam, J., van Der Bent, C., Papapoulos, S.E., et al. Bone morphogenetic proteins stimulate angiogenesis through osteoblast-derived vascular endothelial growth factor A. Endocrinology 143, 1545, 2002.