The effect of timing of mechanical stimulation on proliferation and differentiation of goat bone marrow stem cells cultured on braided PLGA scaffolds.

Bone marrow stromal cells (BMSCs) have been shown to proliferate and produce matrix when seeded onto braided poly(L-lactide/glycolide) acid (PLGA) scaffolds. Mechanical stimulation may be applied to stimulate tissue formation during ligament tissue engineering. This study describes for the first time the effect of constant load on BMSCs seeded onto a braided PLGA scaffold. The seeded scaffolds were subjected to four different loading regimes: Scaffolds were unloaded, loaded during seeding, immediately after seeding, or 2 days after seeding. During the first 5 days, changing the mechanical environment seemed to inhibit proliferation, because cells on scaffolds loaded immediately after seeding or after a 2-day delay, contained fewer cells than on unloaded scaffolds or scaffolds loaded during seeding (p<0.01 for scaffolds loaded after 2 days). During this period, differentiation increased with the period of load applied. After day 5, differences in cell content and collagen production leveled off. After day 11, cell number decreased, whereas collagen production continued to increase. Cell number and differentiation at day 23 were independent of the timing of the mechanical stimulation applied. In conclusion, static load applied to BMSCs cultured on PLGA scaffolds allows for proliferation and differentiation, with loading during seeding yielding the most rapid response. Future research should be aimed at elucidating the biomechanical and biochemical characteristics of tissue formed by BMSCs on PLGA under mechanical stimulation.

[1]  Dennis R. Carter,et al.  Mechanobiology of Skeletal Regeneration , 1998, Clinical orthopaedics and related research.

[2]  J. Goh,et al.  Characterization of a novel polymeric scaffold for potential application in tendon/ligament tissue engineering. , 2006, Tissue engineering.

[3]  Karl Grosh,et al.  Engineering of functional tendon. , 2004, Tissue engineering.

[4]  S. Arnoczky,et al.  Effect of Amplitude and Frequency of Cyclic Tensile Strain on the Inhibition of MMP-1 mRNA Expression in Tendon Cells: An In Vitro Study , 2003, Connective tissue research.

[5]  C. Levene.  Book Review: Treatise on Collagen , 1969 .

[6]  R. Fontana,et al.  Vancomycin-resistant enterococcal colonization and infection in liver transplant candidates and recipients: a prospective surveillance study. , 2006, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[7]  M. Wong,et al.  Cyclic tensile strain and cyclic hydrostatic pressure differentially regulate expression of hypertrophic markers in primary chondrocytes. , 2003, Bone.

[8]  Michael Lavagnino,et al.  Ex vivo static tensile loading inhibits MMP‐1 expression in rat tail tendon cells through a cytoskeletally based mechanotransduction mechanism , 2004, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[9]  Ivan Martin,et al.  Silk matrix for tissue engineered anterior cruciate ligaments. , 2002, Biomaterials.

[10]  T. Kitajima,et al.  Construction of fibroblast–collagen gels with orientated fibrils induced by static or dynamic stress: toward the fabrication of small tendon grafts , 2006, Journal of Artificial Organs.

[11]  M. Bhargava,et al.  Effect of cyclic strain and plating matrix on cell proliferation and integrin expression by ligament fibroblasts , 2006, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[12]  C. Cacou,et al.  Dermal fibroblasts respond to mechanical conditioning in a strain profile dependent manner. , 2003, Biorheology.

[13]  J. Vacanti,et al.  Generation of neo-tendon using synthetic polymers seeded with tenocytes. , 1994, Transplantation proceedings.

[14]  Feng Xu,et al.  In vitro tendon engineering with avian tenocytes and polyglycolic acids: a preliminary report. , 2006, Tissue engineering.

[15]  M. Worsfold,et al.  Microplate assay for the measurement of hydroxyproline in acid-hydrolyzed tissue samples. , 2001, BioTechniques.

[16]  Jack G. Zhou,et al.  Structure and property studies of bioabsorbable poly(glycolide-co-lactide) fiber during processing and in vitro degradation , 2002 .

[17]  E B Hunziker,et al.  Mechanical compression modulates matrix biosynthesis in chondrocyte/agarose culture. , 1995, Journal of cell science.

[18]  J. B. Liesch,et al.  Development of fibroblast-seeded ligament analogs for ACL reconstruction. , 1995, Journal of biomedical materials research.

[19]  Ei Yamamoto,et al.  Effects of static stress on the mechanical properties of cultured collagen fascicles from the rabbit patellar tendon. , 2002, Journal of biomechanical engineering.

[20]  M. Kainer,et al.  Investigation of postoperative allograft-associated infections in patients who underwent musculoskeletal allograft implantation. , 2005, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[21]  David L Kaplan,et al.  Sequential growth factor application in bone marrow stromal cell ligament engineering. , 2005, Tissue engineering.

[22]  K. Sung,et al.  Ligament tissue engineering using synthetic biodegradable fiber scaffolds. , 1999, Tissue engineering.

[23]  W. Willems,et al.  Tissue engineering of ligaments: a comparison of bone marrow stromal cells, anterior cruciate ligament, and skin fibroblasts as cell source. , 2004, Tissue engineering.

[24]  G. N. Ramachandran.  Chemistry of collagen , 1967 .

[25]  Young-Mi Kang,et al.  Nanofiber alignment and direction of mechanical strain affect the ECM production of human ACL fibroblast. , 2005, Biomaterials.

[26]  D. Butler,et al.  In vitro characterization of mesenchymal stem cell-seeded collagen scaffolds for tendon repair: effects of initial seeding density on contraction kinetics. , 2000, Journal of biomedical materials research.

[27]  A J Verbout,et al.  Viable osteogenic cells are obligatory for tissue-engineered ectopic bone formation in goats. , 2003, Tissue engineering.

[28]  Ivan Martin,et al.  The FASEB Journal express article 10.1096/fj.01-0656fje. Published online December 28, 2001. Cell differentiation by mechanical stress , 2022 .

[29]  J. Karlsson,et al.  Donor-site morbidity and anterior knee problems after anterior cruciate ligament reconstruction using autografts. , 2001, Arthroscopy : the journal of arthroscopic & related surgery : official publication of the Arthroscopy Association of North America and the International Arthroscopy Association.

[30]  W. Liu,et al.  Repair of tendon defect with dermal fibroblast engineered tendon in a porcine model. , 2006, Tissue engineering.

[31]  Joseph W Freeman,et al.  Anterior cruciate ligament regeneration using braided biodegradable scaffolds: in vitro optimization studies. , 2005, Biomaterials.

[32]  W. Willems,et al.  Effect of transforming growth factor-beta and growth differentiation factor-5 on proliferation and matrix production by human bone marrow stromal cells cultured on braided poly lactic-co-glycolic acid scaffolds for ligament tissue engineering. , 2007, Tissue engineering.

[33]  C. Lim,et al.  Novel approach to tensile testing of micro- and nanoscale fibers , 2004 .

[34]  R. F. Closkey,et al.  Viability of fibroblast‐seeded ligament analogs after autogenous implantation , 1998, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[35]  J. Kohn,et al.  Preliminary development of a novel resorbable synthetic polymer fiber scaffold for anterior cruciate ligament reconstruction. , 2004, Tissue engineering.

[36]  Joseph W Freeman,et al.  Fiber-based tissue-engineered scaffold for ligament replacement: design considerations and in vitro evaluation. , 2005, Biomaterials.

[37]  D Amiel,et al.  Tendons and ligaments: A morphological and biochemical comparison , 1984, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.