Mechanical Regulation of Bone Regeneration and Vascular Growth in Vivo

Regeneration of large bone defects presents a critical challenge to orthopaedic clinicians as the current treatment strategies are severely limited. Tissue engineering has therefore emerged as a promising alternative to bone grafting techniques. This approach features the delivery of bioactive agents such as stem cells, genes, or proteins using biomaterial delivery systems which together stimulate endogenous repair mechanisms to regenerate the tissue. Because bone is a highly mechanosensitive tissue which responds and adapts dynamically to its mechanical environment, application of mechanical stimuli may enhance endogenous tissue repair. While mechanical loading has been shown to stimulate bone fracture healing, the ability of loading to enhance large bone defect regeneration has not been evaluated.

The goal of this thesis was to evaluate the ability of sustained osteogenic growth factor delivery and functional biomechanical loading to stimulate vascularized repair of large bone defects in a rat segmental defect model. First, we evaluated the hypothesis that the relationship between protein dose and regenerative efficacy depends on delivery system. We determined the dose-response relationship between dose of recombinant human bone morphogenetic protein-2 (rhBMP-2) and bone regeneration in a hybrid alginate-based protein delivery system and compared with the current clinically-used collagen sponge. The hybrid delivery system improved bone formation and reduced the effective dose due to its sustained delivery properties in vivo. Next, we tested the hypothesis that transfer of compressive ambulatory loads during segmental defect repair enhances bone formation and subsequent limb regeneration. We found that delayed application of axial loads enhanced bone regeneration by altering bone formation, tissue differentiation and remodeling, and local strain distribution. Finally, we evaluated the hypothesis that in vivo mechanical loading can enhance neovascular growth to influence bone formation. We found that early mechanical loading disrupted neovascular growth, resulting in impaired bone healing, while delayed loading induced vascular remodeling and enhanced bone formation.

Together, this thesis presents the effects of dose and delivery system on BMPmediated bone regeneration and demonstrates for the first time the effects of in vivo mechanical loading on vascularized regeneration of large bone defects.

Year	Entry
1992	Lanyon LE. The success and failure of the adaptive response to functional load-bearing in averting bone fracture. Bone. 1992;13(2):S17-S21.
1999	Rowley JA, Madlambayan G, Mooney DJ. Alginate hydrogels as synthetic extracellular matrix materials. Biomaterials. January 1999;20(1):45-53.
2007	Rai B, Oest ME, Dupont KM, Ho KH, Teoh SH, Guldberg RE. Combination of platelet‐rich plasma with polycaprolactone‐tricalcium phosphate scaffolds for segmental bone defect repair. J Biomed Mater Res. June 15, 2007;A81(4):888-899.
2004	Simmons CA, Alsberg E, Hsiong S, Kim WJ, Mooney DJ. Dual growth factor delivery and controlled scaffold degradation enhance in vivo bone formation by transplanted bone marrow stromal cells. Bone. August 2004;35(2):562-569.
2006	Sawakami K, Robling AG, Ai M, Pitner ND, Liu D, Warden SJ, Li J, Maye P, Rowe DW, Duncan RL, Warman ML, Turner CH. The Wnt co-receptor LRP5 is essential for skeletal mechanotransduction but not for the anabolic bone response to parathyroid hormone treatment. J Biol Chem. August 18, 2006;281(33):23698-23711.
2006	Robinson JA, Chatterjee-Kishore M, Yaworsky PJ, Cullen DM, Zhao W, Li C, Kharode Y, Sauter L, Babij P, Brown EL, Hill AA, Akhter MP, Johnson ML, Recker RR, Komm BS, Bex FJ. Wnt/β-catenin signaling is a normal physiological response to mechanical loading in bone. J Biol Chem. October 20, 2006;281(42):31720-31728.
2006	Vezeridis PS, Semeins CM, Chen Q, Klein-Nulend J. Osteocytes subjected to pulsating fluid flow regulate osteoblast proliferation and differentiation. Biochem Biophys Res Commun. September 29, 2006;348(3):1082-1088.
1979	Junqueira LCU, Bignolas G, Brentani RR. Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections. Histchem J. 1979;11:447-455.
2002	Kontulainen S, Sievänen H, Kannus P, Pasanen M, Vuori I. Effect of long‐term impact‐loading on mass, size, and estimated strength of humerus and radius of female racquet‐sports players: a peripheral quantitative computed tomography study between young and old starters and controls. J Bone Miner Res. December 2002;17(12):2281-2289.
2002	Deckers MML, van Bezooijen RL, van der Horst G, Hoogendam J, van der Bent C, Papapoulos SE, Löwik CWGM. Bone morphogenetic proteins stimulate angiogenesis through osteoblast-derived vascular endothelial growth factor A. Endocrinology. April 2002;143(4):1545-1553.
1997	Smalt R, Mitchell FT, Howard RL, Chambers TJ. Induction of NO and prostaglandin E₂ in osteoblasts by wall-shear stress but not mechanical strain. Am J Physiol Endocrinol Metab. October 1997;273(4):E751-E758.
1995	Cowin SC, Weinbaum S, Zeng Y. A case for bone canaliculi as the anatomical site of strain generated potentials. J Biomech. November 1995;28(11):1281-1297.
1963	Frost HM. Bone Remodelling Dynamics. Springfield, IL: Charles C. Thomas; 1963.
2011	Kolambkar YM, Dupont KM, Boerckel JD, Huebsch N, Mooney DJ, Hutmacher DW, Guldberg RE. An alginate-based hybrid system for growth factor delivery in the functional repair of large bone defects. Biomaterials. January 2011;32(1):65-74.
2003	Cartmell SH, Porter BD, García AJ, Guldberg RE. Effects of medium perfusion rate on cell-seeded three-dimensional bone constructs in vitro. Tissue Eng. December 2003;9(6):1197-1203.
2001	Ryder KD, Duncan RL. Parathyroid hormone enhances fluid shear-induced [Ca²⁺]_i signaling in osteoblastic cells through activation of mechanosensitive and voltage-sensitive Ca²⁺ channels. J Bone Miner Res. February 2001;16(2):240-248.
1999	Gerber H-P, Vu TH, Ryan AM, Kowalski J, Werb Z, Ferrara N. VEGF couples hypertrophic cartilage remodeling, ossification and angiogenesis during endochondral bone formation. Nat Med. June 1999;5(6):623-628.
2006	Swiontkowski MF, Aro HT, Donell S, Esterhai JL, Goulet J, Jones A, Kregor PJ, Nordsletten L, Paiement G, Patel A. Recombinant human bone morphogenetic protein-2 in open tibial fractures: a subgroup analysis of data combined from two prospective randomized studies. J Bone Joint Surg. June 2006;88A(6):1258-1265.
2004	Knothe Tate ML, Adamson JR, Tami AE, Bauer TW. The osteocyte. Int J Biochem Cell Biol. January 2004;36(1):1-8.
2007	Malone AMD, Anderson CT, Tummala P, Kwon RY, Johnston TR, Stearns T, Jacobs CR. Primary cilia mediate mechanosensing in bone cells by a calcium-independent mechanism. Proc Natl Acad Sci USA. August 14, 2007;104(33):13325-13330.
1997	Jaiswal N, Haynesworth SE, Caplan AI, Bruder SP. Osteogenic differentiation of purified, culture‐expanded human mesenchymal stem cells in vitro. J Cell Biochem. February 1997;64(2):295-312.
1997	Owan I, Burr DB, Turner CH, Qiu J, Tu Y, Onyia JE, Duncan RL. Mechanotransduction in bone: osteoblasts are more responsive to fluid forces than mechanical strain. Am J Physiol Cell Physiol. September 1997;273(3):C810-C815.
1998	Claes LE, Heigele CA, Neidlinger-Wilke C, Kaspar D, Seidl W, Margevicius KJ, Augat P. Effects of mechanical factors on the fracture healing process. Clin Orthop Relat Res. October 1998;355(suppl):S132-S147.
1999	Claes LE, Heigele CA. Magnitudes of local stress and strain along bony surfaces predict the course and type of fracture healing. J Biomech. March 1999;32(3):255-266.
1995	Turner CH, Owan I, Takano Y. Mechanotransduction in bone: role of strain rate. Am J Physiol Endocrinol Metab. September 1995;269(3):E438-E442.
1979	Perren SM. Physical and biological aspects of fracture healing with special reference to internal fixation. Clin Orthop Relat Res. January–February 1979;138:175-196.
2006	Rezwan K, Chen QZ, Blaker JJ, Boccaccini AR. Biodegradable and bioactive porous polymer/inorganic composite scaffolds for bone tissue engineering. Biomaterials. June 2006;27(18):3413-3431.
2006	Xie L, Jacobson JM, Choi ES, Busa B, Donahue LR, Miller LM, Rubin CT, Judex S. Low-level mechanical vibrations can influence bone resorption and bone formation in the growing skeleton. Bone. November 2006;39(5):1059-1066.
1998	Carter DR, Beaupré GS, Giori NJ, Helms JA. Mechanobiology of skeletal regeneration. Clin Orthop Relat Res. October 1998;355(suppl):S41-S55.
1992	van der Plas A, Nijweide PJ. Isolation and purification of osteocytes. J Bone Miner Res. April 1992;7(4):389-396.
1977	Piekarski K, Munro M. Transport mechanism operating between blood supply and osteocytes in long bones. Nature. September 1, 1977;270(5623):80-82.
2006	Rubin J, Rubin C, Jacobs CR. Molecular pathways mediating mechanical signaling in bone. Gene. February 15, 2006;367:1-16.
1988	Wozney JM, Rosen V, Celeste AJ, Mitsock LM, Whitters MJ, Kriz RW, Hewick RM, Wang EA. Novel regulators of bone formation: molecular clones and activities. Science. December 16, 1988;242(4885):1528-1534.
2002	Qin Y-X, Lin W, Rubin C. The pathway of bone fluid flow as defined by in vivo intramedullary pressure and streaming potential measurements. Ann Biomed Eng. May 2002;30(5):693-702.
2008	Bonewald LF, Johnson ML. Osteocytes, mechanosensing and Wnt signaling. Bone. April 2008;42(4):606-615.
1987	Frost HM. Bone "mass" and the "mechanostat": a proposal. Anat Rec. September 1987;219(1):1-9.
1965	Urist MR. Bone: formation by autoinduction. Science. November 12, 1965;150(3698):893-899.
2007	Oest ME, Dupont KM, Kong H, Mooney DJ, Guldberg RE. Quantitative assessment of scaffold and growth factor‐mediated repair of critically sized bone defects. J Orthop Res. July 2007;25(7):941-950.
2009	Boerckel JD, Dupont KM, Kolambkar YM, Lin ASP, Guldberg RE. In vivo model for evaluating the effects of mechanical stimulation on tissue-engineered bone repair. J Biomech Eng. August 2009;131(8):084502.
1995	Duncan RL, Turner CH. Mechanotransduction and the functional response of bone to mechanical strain. Calcif Tiss Int. December 1995;57(5):344-358.
1984	Lanyon LE, Rubin CT. Static vs dynamic loads as an influence on bone remodelling. J Biomech. 1984;17(12):897-905.
2000	Haapasalo H, Kontulainen S, Sievänen H, Kannus P, Järvinen M, Vuori I. Exercise-induced bone gain is due to enlargement in bone size without a change in volumetric bone density: a peripheral quantitative computed tomography study of the upper arms of male tennis players. Bone. September 2000;27(3):351-357.
2010	Dupont KM, Sharma K, Stevens HY, Boerckel JD, García AJ, Guldberg RE. Human stem cell delivery for treatment of large segmental bone defects. Proc Natl Acad Sci USA. February 23, 2010;107(8):3305-3310.
2001	Sikavitsas VI, Temenoff JS, Mikos AG. Biomaterials and bone mechanotransduction. Biomaterials. October 2001;22(19):2581-2593.
1994	Turner CH, Forwood MR, Rho J, Yoshikawa T. Mechanical loading thresholds for lamellar and woven bone formation. J Bone Miner Res. January 1994;9(1):87-97.
1997	Guldberg RE, Caldwell NJ, Guo XE, Goulet RW, Hollister SJ, Goldstein SA. Mechanical stimulation of tissue repair in the hydraulic bone chamber. J Bone Miner Res. August 1997;12(8):1295-1302.
2003	Young MF. Bone matrix proteins: their function, regulation, and relationship to osteoporosis. Osteoporos Int. March 2003;14(suppl 3):S35-S42.
1984	Rubin CT, Lanyon LE. Regulation of bone formation by applied dynamic loads. J Bone Joint Surg. March 1984;66A(3):397-402.
1995	Klein-Nulend J, van der Plas A, Semeins CM, Ajubi NE, Frangos JA, Nijweide PJ, Burger EH. Sensitivity of osteocytes to biomechanical stress in vitro. FASEB J. March 1995;9(5):441-445.
2011	Price C, Zhou X, Li W, Wang L. Real‐time measurement of solute transport within the lacunar‐canalicular system of mechanically loaded bone: direct evidence for load‐induced fluid flow. J Bone Miner Res. February 2011;26(2):277-285.
1987	Carter DR, Fyhrie DP, Whalen RT. Trabecular bone density and loading history: regulation of connective tissue biology by mechanical energy. J Biomech. 1987;20(8):785-794.
2004	Duvall CL, Taylor WR, Weiss D, Guldberg RE. Quantitative microcomputed tomography analysis of collateral vessel development after ischemic injury. Am J Physiol Heart Circ Physiol. July 2004;287(1):H302-H310.
2001	Hausman MR, Schaffler MB, Majeska RJ. Prevention of fracture healing in rats by an inhibitor of angiogenesis. Bone. December 2001;29(6):560-564.
1994	Weinbaum S, Cowin SC, Zeng Y. A model for the excitation of osteocytes by mechanical loading-induced bone fluid shear stresses. J Biomech. March 1994;27(3):339-360.
2002	Lieberman JR, Daluiski A, Einhorn TA. The role of growth factors in the repair of bone: biology and clinical applications. J Bone Joint Surg. June 2002;84A(6):1032-1044.
2002	Street J, Bao M, deGuzman L, Bunting S, Peale FV Jr, Ferrara N, Steinmetz H, Hoeffel J, Cleland JL, Daugherty A, van Bruggen N, Redmond HP, Carano RAD, Filvaroff EH. Vascular endothelial growth factor stimulates bone repair by promoting angiogenesis and bone turnover. Proc Natl Acad Sci USA. July 23, 2002;99(15):9656-9661.
2007	Cowin SC, Doty SB. Tissue Mechanics. Boca Raton, FL: Springer; 2007.
1985	Goodship AE, Kenwright J. The influence of induced micromovement upon the healing of experimental tibial fractures. J Bone Joint Surg. 1985;67B(4):650-655.
2003	Gerstenfeld LC, Cullinane DM, Barnes GL, Graves DT, Einhorn TA. Fracture healing as a post‐natal developmental process: molecular, spatial, and temporal aspects of its regulation. J Cell Biochem. April 2003;88(5):873-884.
1977	Jones HH, Priest JD, Hayes WC, Tichenor CC, Nagel DA. Humeral hypertrophy in response to exercise. J Bone Joint Surg. March 1977;59A(2):204-208.
2001	Bakker AD, Soejima K, Klein-Nulend J, Burger EH. The production of nitric oxide and prostaglandin E₂ by primary bone cells is shear stress dependent. J Biomech. May 2001;34(5):671-677.
1996	Carter DR, Van der Meulen MCH, Beaupré GS. Mechanical factors in bone growth and development. Bone. January 1996;18(1)(suppl 1):S5-S10.
1998	Turner CH, Pavalko FM. Mechanotransduction and functional response of the skeleton to physical stress: the mechanisms and mechanics of bone adaptation. J Orthop Sci. November 1998;3(6):346-355.
1998	Turner CH. Three rules for bone adaptation to mechanical stimuli. Bone. November 1998;23(5):399-407.
2008	Shapiro F. Bone development and its relation to fracture repair: the role of mesenchymal osteoblasts and surface osteoblasts. Eur Cell Mater. 2008;15:53-76.
1998	Knothe Tate ML, Knothe U, Niederer P. Experimental elucidation of mechanical load-induced fluid flow and its potential role in bone metabolism and functional adaptation. Am J Med Sci. September 1998;316(3):189-195.
1998	Mosley JR, Lanyon LE. Strain rate as a controlling influence on adaptive modeling in response to dynamic loading of the ulna in growing male rats. Bone. October 1998;23(4):313-318.
1985	Rubin CT, Lanyon LE. Regulation of bone mass by mechanical strain magnitude. Calcif Tiss Int. 1985;37(4):411-417.
2001	Cowin SC, ed. Bone Mechanics Handbook. 2nd ed. Boca Raton, FL: CRC Press; 2001.
2006	Gilsanz V, Wren TAL, Sanchez M, Dorey F, Judex S, Rubin C. Low‐level, high‐frequency mechanical signals enhance musculoskeletal development of young women with low bmd. J Bone Miner Res. September 2006;21(9):1464-1474.
2004	Han Y, Cowin SC, Schaffler MB, Weinbaum S. Mechanotransduction and strain amplification in osteocyte cell processes. Proc Natl Acad Sci USA. November 23, 2004;101(47):16689-16694.
1995	Burger EH, Klein-Nulend J, Van Der Plas A, Nijweide PJ. Function of osteocytes in bone: their role in mechanotransduction. J Nutr. July 1995;125(7)(suppl):S2020-S2023.
2006	Robling AG, Castillo AB, Turner CH. Biomechanical and molecular regulation of bone remodeling. Annu Rev Biomed Eng. 2006;8:455-498.
1997	Mullender MG, Huiskes R. Osteocytes and bone lining cells: which are the best candidates for mechano-sensors in cancellous bone? Bone. June 1997;20(6):527-532.

Year

Entry

1992

Lanyon LE. The success and failure of the adaptive response to functional load-bearing in averting bone fracture. Bone. 1992;13(2):S17-S21.

1999

Rowley JA, Madlambayan G, Mooney DJ. Alginate hydrogels as synthetic extracellular matrix materials. Biomaterials. January 1999;20(1):45-53.

2007

Rai B, Oest ME, Dupont KM, Ho KH, Teoh SH, Guldberg RE. Combination of platelet‐rich plasma with polycaprolactone‐tricalcium phosphate scaffolds for segmental bone defect repair. J Biomed Mater Res. June 15, 2007;A81(4):888-899.

2004

Simmons CA, Alsberg E, Hsiong S, Kim WJ, Mooney DJ. Dual growth factor delivery and controlled scaffold degradation enhance in vivo bone formation by transplanted bone marrow stromal cells. Bone. August 2004;35(2):562-569.

2006

Sawakami K, Robling AG, Ai M, Pitner ND, Liu D, Warden SJ, Li J, Maye P, Rowe DW, Duncan RL, Warman ML, Turner CH. The Wnt co-receptor LRP5 is essential for skeletal mechanotransduction but not for the anabolic bone response to parathyroid hormone treatment. J Biol Chem. August 18, 2006;281(33):23698-23711.

2006

Robinson JA, Chatterjee-Kishore M, Yaworsky PJ, Cullen DM, Zhao W, Li C, Kharode Y, Sauter L, Babij P, Brown EL, Hill AA, Akhter MP, Johnson ML, Recker RR, Komm BS, Bex FJ. Wnt/β-catenin signaling is a normal physiological response to mechanical loading in bone. J Biol Chem. October 20, 2006;281(42):31720-31728.

2006

Vezeridis PS, Semeins CM, Chen Q, Klein-Nulend J. Osteocytes subjected to pulsating fluid flow regulate osteoblast proliferation and differentiation. Biochem Biophys Res Commun. September 29, 2006;348(3):1082-1088.

1979

Junqueira LCU, Bignolas G, Brentani RR. Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections. Histchem J. 1979;11:447-455.

2002

Kontulainen S, Sievänen H, Kannus P, Pasanen M, Vuori I. Effect of long‐term impact‐loading on mass, size, and estimated strength of humerus and radius of female racquet‐sports players: a peripheral quantitative computed tomography study between young and old starters and controls. J Bone Miner Res. December 2002;17(12):2281-2289.

2002

Deckers MML, van Bezooijen RL, van der Horst G, Hoogendam J, van der Bent C, Papapoulos SE, Löwik CWGM. Bone morphogenetic proteins stimulate angiogenesis through osteoblast-derived vascular endothelial growth factor A. Endocrinology. April 2002;143(4):1545-1553.

1997

Smalt R, Mitchell FT, Howard RL, Chambers TJ. Induction of NO and prostaglandin E₂ in osteoblasts by wall-shear stress but not mechanical strain. Am J Physiol Endocrinol Metab. October 1997;273(4):E751-E758.

1995

Cowin SC, Weinbaum S, Zeng Y. A case for bone canaliculi as the anatomical site of strain generated potentials. J Biomech. November 1995;28(11):1281-1297.

1963

Frost HM. Bone Remodelling Dynamics. Springfield, IL: Charles C. Thomas; 1963.

2011

Kolambkar YM, Dupont KM, Boerckel JD, Huebsch N, Mooney DJ, Hutmacher DW, Guldberg RE. An alginate-based hybrid system for growth factor delivery in the functional repair of large bone defects. Biomaterials. January 2011;32(1):65-74.

2003

Cartmell SH, Porter BD, García AJ, Guldberg RE. Effects of medium perfusion rate on cell-seeded three-dimensional bone constructs in vitro. Tissue Eng. December 2003;9(6):1197-1203.

2001

Ryder KD, Duncan RL. Parathyroid hormone enhances fluid shear-induced [Ca²⁺]_i signaling in osteoblastic cells through activation of mechanosensitive and voltage-sensitive Ca²⁺ channels. J Bone Miner Res. February 2001;16(2):240-248.

1999

Gerber H-P, Vu TH, Ryan AM, Kowalski J, Werb Z, Ferrara N. VEGF couples hypertrophic cartilage remodeling, ossification and angiogenesis during endochondral bone formation. Nat Med. June 1999;5(6):623-628.

2006

Swiontkowski MF, Aro HT, Donell S, Esterhai JL, Goulet J, Jones A, Kregor PJ, Nordsletten L, Paiement G, Patel A. Recombinant human bone morphogenetic protein-2 in open tibial fractures: a subgroup analysis of data combined from two prospective randomized studies. J Bone Joint Surg. June 2006;88A(6):1258-1265.

2004

Knothe Tate ML, Adamson JR, Tami AE, Bauer TW. The osteocyte. Int J Biochem Cell Biol. January 2004;36(1):1-8.

2007

Malone AMD, Anderson CT, Tummala P, Kwon RY, Johnston TR, Stearns T, Jacobs CR. Primary cilia mediate mechanosensing in bone cells by a calcium-independent mechanism. Proc Natl Acad Sci USA. August 14, 2007;104(33):13325-13330.

1997

Jaiswal N, Haynesworth SE, Caplan AI, Bruder SP. Osteogenic differentiation of purified, culture‐expanded human mesenchymal stem cells in vitro. J Cell Biochem. February 1997;64(2):295-312.

1997

Owan I, Burr DB, Turner CH, Qiu J, Tu Y, Onyia JE, Duncan RL. Mechanotransduction in bone: osteoblasts are more responsive to fluid forces than mechanical strain. Am J Physiol Cell Physiol. September 1997;273(3):C810-C815.

1998

Claes LE, Heigele CA, Neidlinger-Wilke C, Kaspar D, Seidl W, Margevicius KJ, Augat P. Effects of mechanical factors on the fracture healing process. Clin Orthop Relat Res. October 1998;355(suppl):S132-S147.

1999

Claes LE, Heigele CA. Magnitudes of local stress and strain along bony surfaces predict the course and type of fracture healing. J Biomech. March 1999;32(3):255-266.

1995

Turner CH, Owan I, Takano Y. Mechanotransduction in bone: role of strain rate. Am J Physiol Endocrinol Metab. September 1995;269(3):E438-E442.

1979

Perren SM. Physical and biological aspects of fracture healing with special reference to internal fixation. Clin Orthop Relat Res. January–February 1979;138:175-196.

2006

Rezwan K, Chen QZ, Blaker JJ, Boccaccini AR. Biodegradable and bioactive porous polymer/inorganic composite scaffolds for bone tissue engineering. Biomaterials. June 2006;27(18):3413-3431.

2006

Xie L, Jacobson JM, Choi ES, Busa B, Donahue LR, Miller LM, Rubin CT, Judex S. Low-level mechanical vibrations can influence bone resorption and bone formation in the growing skeleton. Bone. November 2006;39(5):1059-1066.

1998

Carter DR, Beaupré GS, Giori NJ, Helms JA. Mechanobiology of skeletal regeneration. Clin Orthop Relat Res. October 1998;355(suppl):S41-S55.

1992

van der Plas A, Nijweide PJ. Isolation and purification of osteocytes. J Bone Miner Res. April 1992;7(4):389-396.

1977

Piekarski K, Munro M. Transport mechanism operating between blood supply and osteocytes in long bones. Nature. September 1, 1977;270(5623):80-82.

2006

Rubin J, Rubin C, Jacobs CR. Molecular pathways mediating mechanical signaling in bone. Gene. February 15, 2006;367:1-16.

1988

Wozney JM, Rosen V, Celeste AJ, Mitsock LM, Whitters MJ, Kriz RW, Hewick RM, Wang EA. Novel regulators of bone formation: molecular clones and activities. Science. December 16, 1988;242(4885):1528-1534.

2002

Qin Y-X, Lin W, Rubin C. The pathway of bone fluid flow as defined by in vivo intramedullary pressure and streaming potential measurements. Ann Biomed Eng. May 2002;30(5):693-702.

2008

Bonewald LF, Johnson ML. Osteocytes, mechanosensing and Wnt signaling. Bone. April 2008;42(4):606-615.

1987

Frost HM. Bone "mass" and the "mechanostat": a proposal. Anat Rec. September 1987;219(1):1-9.

1965

Urist MR. Bone: formation by autoinduction. Science. November 12, 1965;150(3698):893-899.

2007

Oest ME, Dupont KM, Kong H, Mooney DJ, Guldberg RE. Quantitative assessment of scaffold and growth factor‐mediated repair of critically sized bone defects. J Orthop Res. July 2007;25(7):941-950.

2009

Boerckel JD, Dupont KM, Kolambkar YM, Lin ASP, Guldberg RE. In vivo model for evaluating the effects of mechanical stimulation on tissue-engineered bone repair. J Biomech Eng. August 2009;131(8):084502.

1995

Duncan RL, Turner CH. Mechanotransduction and the functional response of bone to mechanical strain. Calcif Tiss Int. December 1995;57(5):344-358.

1984

Lanyon LE, Rubin CT. Static vs dynamic loads as an influence on bone remodelling. J Biomech. 1984;17(12):897-905.

2000

Haapasalo H, Kontulainen S, Sievänen H, Kannus P, Järvinen M, Vuori I. Exercise-induced bone gain is due to enlargement in bone size without a change in volumetric bone density: a peripheral quantitative computed tomography study of the upper arms of male tennis players. Bone. September 2000;27(3):351-357.

2010

Dupont KM, Sharma K, Stevens HY, Boerckel JD, García AJ, Guldberg RE. Human stem cell delivery for treatment of large segmental bone defects. Proc Natl Acad Sci USA. February 23, 2010;107(8):3305-3310.

2001

Sikavitsas VI, Temenoff JS, Mikos AG. Biomaterials and bone mechanotransduction. Biomaterials. October 2001;22(19):2581-2593.

1994

Turner CH, Forwood MR, Rho J, Yoshikawa T. Mechanical loading thresholds for lamellar and woven bone formation. J Bone Miner Res. January 1994;9(1):87-97.

1997

Guldberg RE, Caldwell NJ, Guo XE, Goulet RW, Hollister SJ, Goldstein SA. Mechanical stimulation of tissue repair in the hydraulic bone chamber. J Bone Miner Res. August 1997;12(8):1295-1302.

2003

Young MF. Bone matrix proteins: their function, regulation, and relationship to osteoporosis. Osteoporos Int. March 2003;14(suppl 3):S35-S42.

1984

Rubin CT, Lanyon LE. Regulation of bone formation by applied dynamic loads. J Bone Joint Surg. March 1984;66A(3):397-402.

1995

Klein-Nulend J, van der Plas A, Semeins CM, Ajubi NE, Frangos JA, Nijweide PJ, Burger EH. Sensitivity of osteocytes to biomechanical stress in vitro. FASEB J. March 1995;9(5):441-445.

2011

Price C, Zhou X, Li W, Wang L. Real‐time measurement of solute transport within the lacunar‐canalicular system of mechanically loaded bone: direct evidence for load‐induced fluid flow. J Bone Miner Res. February 2011;26(2):277-285.

1987

Carter DR, Fyhrie DP, Whalen RT. Trabecular bone density and loading history: regulation of connective tissue biology by mechanical energy. J Biomech. 1987;20(8):785-794.

2004

Duvall CL, Taylor WR, Weiss D, Guldberg RE. Quantitative microcomputed tomography analysis of collateral vessel development after ischemic injury. Am J Physiol Heart Circ Physiol. July 2004;287(1):H302-H310.

2001

Hausman MR, Schaffler MB, Majeska RJ. Prevention of fracture healing in rats by an inhibitor of angiogenesis. Bone. December 2001;29(6):560-564.

1994

Weinbaum S, Cowin SC, Zeng Y. A model for the excitation of osteocytes by mechanical loading-induced bone fluid shear stresses. J Biomech. March 1994;27(3):339-360.

2002

Lieberman JR, Daluiski A, Einhorn TA. The role of growth factors in the repair of bone: biology and clinical applications. J Bone Joint Surg. June 2002;84A(6):1032-1044.

2002

Street J, Bao M, deGuzman L, Bunting S, Peale FV Jr, Ferrara N, Steinmetz H, Hoeffel J, Cleland JL, Daugherty A, van Bruggen N, Redmond HP, Carano RAD, Filvaroff EH. Vascular endothelial growth factor stimulates bone repair by promoting angiogenesis and bone turnover. Proc Natl Acad Sci USA. July 23, 2002;99(15):9656-9661.

2007

Cowin SC, Doty SB. Tissue Mechanics. Boca Raton, FL: Springer; 2007.

1985

Goodship AE, Kenwright J. The influence of induced micromovement upon the healing of experimental tibial fractures. J Bone Joint Surg. 1985;67B(4):650-655.

2003

Gerstenfeld LC, Cullinane DM, Barnes GL, Graves DT, Einhorn TA. Fracture healing as a post‐natal developmental process: molecular, spatial, and temporal aspects of its regulation. J Cell Biochem. April 2003;88(5):873-884.

1977

Jones HH, Priest JD, Hayes WC, Tichenor CC, Nagel DA. Humeral hypertrophy in response to exercise. J Bone Joint Surg. March 1977;59A(2):204-208.

2001

Bakker AD, Soejima K, Klein-Nulend J, Burger EH. The production of nitric oxide and prostaglandin E₂ by primary bone cells is shear stress dependent. J Biomech. May 2001;34(5):671-677.

1996

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Year	Entry
2013	Willett NJ, Li M-TA, Uhrig BA, Boerckel JD, Huebsch N, Lundgren TL, Warren GL, Guldberg RE. Attenuated human bone morphogenetic protein-2–mediated bone regeneration in a rat model of composite bone and muscle injury. Tissue Eng Part C Methods. April 2013;19(4):316-325.
2015	Priddy LB. Biomaterial Strategies for Improved Bone Healing With Bone Morphogenetic Protein-2 Delivery [PhD thesis]. Atlanta, GA: Georgia Institute of Technology; August 2015.

Year

Entry

2013

Willett NJ, Li M-TA, Uhrig BA, Boerckel JD, Huebsch N, Lundgren TL, Warren GL, Guldberg RE. Attenuated human bone morphogenetic protein-2–mediated bone regeneration in a rat model of composite bone and muscle injury. Tissue Eng Part C Methods. April 2013;19(4):316-325.

2015

Priddy LB. Biomaterial Strategies for Improved Bone Healing With Bone Morphogenetic Protein-2 Delivery [PhD thesis]. Atlanta, GA: Georgia Institute of Technology; August 2015.