Distinctive degradation behaviors of electrospun polyglycolide, poly(DL-lactide-co-glycolide), and poly(L-lactide-co-epsilon-caprolactone) nanofibers cultured with/without porcine smooth muscle cells.
Biodegradable nanofibers have become a popular candidate for tissue engineering scaffolds because of their biomimetic structure that physically resembles the extracellular matrix. For certain tissue regeneration applications, prolonged in vitro culture time for cellular reorganization and tissue remodeling may be required. Therefore, extensive understanding of cellular effects on scaffold degradation is needed. There are only few studies on the degradation of nanofibers, and also the studies on degradation throughout cell culture are rare. In this study, polyglycolide (PGA), poly(DL-lactide-co-glycolide) (PLGA) and poly(L-lactide-co-epsilon-caprolactone) [P(LLA-CL)] were electrospun into nanofibrous meshes. The nanofibers were cultured with porcine smooth muscle cells for up to 3 months to evaluate their degradation behavior and cellular response. The results showed that the degradation rates are in the order of PGA >> PLGA > P(LLA-CL). PGA nanofibers degraded in 3 weeks and supported cell growth only in the first few days. PLGA nanofiber scaffolds facilitated cell growth during the first 30 days after seeding, but cell growth was slow thereafter. P(LLA-CL) nanofibers facilitated long-term (1-3 months) cell growth. mRNA quantification using real-time polymerase chain reaction revealed that some smooth muscle cell markers (alpha-actinin and calponin) and extracellular matrix genes (collagen and integrin) seemed to be downregulated with increased cell culture time. Cell culture significantly increased the degradation rate of PGA nanofibers, whereas the effect on PLGA and P(LLA-CL) nanofibers was limited. We found that the molecular weight of P(LLA-CL) and PLGA nanofibers decreased linearly for up to 100 days. Half lives of PLGA and P(LLA-CL) nanofibers were shown to be 80 and 110 days, respectively. In summary, this is the first study to our knowledge to evaluate long-term polymeric nanofiber degradation in vitro with cell culture. Cell culture accelerated the nanofibrous scaffold degradation to a limited extent. P(LLA-CL) nanofibers could be a good choice as scaffolds for long-term smooth muscle cell culture.
Division of Bioengineering, National University of Singapore, Singapore, Singapore.
Yixiang
Y
Dong
Thomas
T
Yong
Susan
S
Liao
Casey K
CK
Chan
Molly M
MM
Stevens
Seeram
S
Ramakrishna
D020341:Absorbable Implants; D000818:Animals; D040761:Biomimetic Materials; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D005109:Extracellular Matrix; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis; D005786:Gene Expression Regulation; D019344:Lactic Acid; D032389:Myocytes, Smooth Muscle / Q000378:metabolism / Q000648:ultrastructure*; D057139:Nanofibers; D011091:Polyesters; D011100:Polyglycolic Acid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D013552:Swine; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods
D016326:Extracellular Matrix Proteins; D011091:Polyesters; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; C062968:lactide-caprolactone copolymer