TY - JOUR
T1 - Design and characterization of microporous hyaluronic acid hydrogels for in vitro gene transfer to mMSCs
AU - Tokatlian, Talar
AU - Cam, Cynthia
AU - Siegman, Shayne N.
AU - Lei, Yuguo
AU - Segura, Tatiana
N1 - Funding Information:
The authors would like to thank James Dorman for his help with SEM imaging, Shiva Gojgini for her assistance in developing the DNA radio labeling protocol, as well as Jonathan Lam and Maha Rahim for their help modifying hyaluronic acid. The authors acknowledge the NIH ( R01HL110592 ), and NSF ( CAREER 0747539 ), for funding this work. T. Tokatlian also acknowledges the NIH Biotechnology Training Grant ( T32GM067555 ) for funding.
PY - 2012/11
Y1 - 2012/11
N2 - The effective and sustained delivery of DNA locally could increase the applicability of gene therapy in tissue regeneration and therapeutic angiogenesis. One promising approach is to use porous hydrogel scaffolds to encapsulate and deliver nucleotides in the form of nanoparticles to the affected sites. We have designed and characterized microporous (μ-pore) hyaluronic acid hydrogels which allow for effective cell seeding in vitro post-scaffold fabrication and allow for cell spreading and proliferation without requiring high levels of degradation. These factors, coupled with high loading efficiency of DNA polyplexes using a previously developed caged nanoparticle encapsulation (CnE) technique, then allowed for long-term sustained transfection and transgene expression of incorporated mMSCs. In this study, we examined the effect of pore size on gene transfer efficiency and the kinetics of transgene expression. For all investigated pore sizes (30, 60, and 100 μm), encapsulated DNA polyplexes were released steadily, starting by day 4 for up to 10 days. Likewise, transgene expression was sustained over this period, although significant differences between different pore sizes were not observed. Cell viability was also shown to remain high over time, even in the presence of high concentrations of DNA polyplexes. The knowledge acquired through this in vitro model can be utilized to design and better predict scaffold-mediated gene delivery for local gene therapy in an in vivo model where host cells infiltrate the scaffold over time.
AB - The effective and sustained delivery of DNA locally could increase the applicability of gene therapy in tissue regeneration and therapeutic angiogenesis. One promising approach is to use porous hydrogel scaffolds to encapsulate and deliver nucleotides in the form of nanoparticles to the affected sites. We have designed and characterized microporous (μ-pore) hyaluronic acid hydrogels which allow for effective cell seeding in vitro post-scaffold fabrication and allow for cell spreading and proliferation without requiring high levels of degradation. These factors, coupled with high loading efficiency of DNA polyplexes using a previously developed caged nanoparticle encapsulation (CnE) technique, then allowed for long-term sustained transfection and transgene expression of incorporated mMSCs. In this study, we examined the effect of pore size on gene transfer efficiency and the kinetics of transgene expression. For all investigated pore sizes (30, 60, and 100 μm), encapsulated DNA polyplexes were released steadily, starting by day 4 for up to 10 days. Likewise, transgene expression was sustained over this period, although significant differences between different pore sizes were not observed. Cell viability was also shown to remain high over time, even in the presence of high concentrations of DNA polyplexes. The knowledge acquired through this in vitro model can be utilized to design and better predict scaffold-mediated gene delivery for local gene therapy in an in vivo model where host cells infiltrate the scaffold over time.
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U2 - 10.1016/j.actbio.2012.07.014
DO - 10.1016/j.actbio.2012.07.014
M3 - Article
C2 - 22820309
AN - SCOPUS:84873940453
SN - 1742-7061
VL - 8
SP - 3921
EP - 3931
JO - Acta Biomaterialia
JF - Acta Biomaterialia
IS - 11
ER -