Fahriye Duygu ÇETİNKAYA, Sevil KÖSE, Fatima KAYA AERTS, Emir Bakır DENKBAŞ, Petek KORKUSUZ

Evaluation of biocompatibility of random or aligned electrospun polyhydroxybutyrate scaffolds combined with human mesenchymal stem cells

Polyhydroxybutyrate (PHB) is a polymer used to restore tissues or regenerate bones. However, its compatibility with human bone marrow-derived mesenchymal stem cells (MSCs) has not been examined. PHB membranes of random (r-PHB) or aligned (a-PHB) electrospun nanofibers that generate bone and spinal axon scaffolds were combined with human MSCs. The adhesion and proliferation of cells on these membranes were examined. The orientation of cells on PHB membranes was analyzed by confocal microscopy and scanning electron microscopy (SEM). The MSCs maintained their characteristic properties on the membranes, adhered to the membranes, and preserved their viability. The cell morphology was different when they were grown on differentially designed matrices. Cells expanded on the a-PHB membrane and showed fibroid-like morphology. Conversely, cells interacting with the r-PHB membrane were located homogeneously and demonstrated polygonal morphology. The adhesion and proliferation of human MSCs was higher on the a-PHB membrane than on the randomly oriented one. Fiber orientation influenced the phenotype and biological behavior of human MSCs. This property may be useful in the selection of specifically designed scaffolds for the desired tasks. The results of this preliminary study indicated that PHB membranes designed for bone or nerve tissue engineering are compatible with human MSCs.

PDF

___

Asran AS, Razghandi K, Aggarwal N, Michler GH, Groth T (2010). Nanofibers from blends of polyvinyl alcohol and polyhydroxy butyrate as potential scaffold material for tissue engineering of skin. Biomacromolecules 11: 3413–3421.
Chen GQ, Wu Q (2005). The application of polyhydroxyalkanoates as tissue engineering materials. Biomaterials 26: 6565–6578.
Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Prockop DJ, Horwitz E (2006). Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8: 315–317.
Engelberg I, Kohn J (1991). Physico-mechanical properties of degradable polymers used in medical applications: a comparative study. Biomaterials 12: 292–304.
Freier T, Kunze C, Nischan C, Kramer S, Sternberg K, Sass M, Hopt UT, Schmitz KP (2002). In vitro and in vivo degradation studies for development of a biodegradable patch based on poly(3-hydroxybutyrate). Biomaterials 23: 2649–2657.
Galego N, Rosza C, Sanchez R, Fung J, Vazquez A, Tomas J (2000). Characterization and application of poly(β-hydroxyalkanoates) family as composite biomaterials. Polym Test 19: 485–492.
Hu YJ, Wei X, Zhao W, Liu YS, Chen GQ (2009). Biocompatibility of poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-3- hydroxyhexanoate) with bone marrow mesenchymal stem cells. Acta Biomater 5: 1115–1125.
Khorasania MT, Mirmohammadia SA, Irania S (2011). Polyhydroxybutyrate (PHB) scaffolds as a model for nerve tissue engineering application: fabrication and in vitro assay. Int J Polym Mater 60: 562–575.
Köse GT, Kenar H, Hasirci N, Hasirci V (2003). Macroporous poly(3-hydroxybutyrate-co-3-hydroxyvalerate) matrices for bone tissue engineering. Biomaterials 24: 1949–1958.
Köse GT, Korkusuz F, Korkusuz P, Purali N, Ozkul A, Hasirci V (2003). Bone generation on PHBV matrices: an in vitro study. Biomaterials 24: 4999–5007.
Li M, Mondrinos MJ, Gandhi MR, Ko FK, Weiss AS, Lelkes PI (2005). Electrospun protein fibers as matrices for tissue engineering. Biomaterials 26: 5999–6008.
Li XT, Zhang Y, Chen GQ (2008). Nanofibrous polyhydroxyalkanoate matrices as cell growth supporting materials. Biomaterials 29: 3720–3728.
Lutolf MP, Hubbell JA (2005). Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering. Nat Biotechnol 23: 47–55.
Masaeli E, Wieringa PA, Morshed M, Nasr-Esfahani MH, Sadri S, van Blitterswijk CA, Moroni L (2014). Peptide functionalized polyhydroxyalkanoate nanofibrous scaffolds enhance Schwann cells activity. Nanomedicine-UK 10: 1559–1569.
McLane JS, Schaub NJ, Gilbert RJ, Ligon LA (2013). Electrospun nanofiber scaffolds for investigating cell-matrix adhesion. Method Mol Cell Biol 1046: 371–388.
Misra SK, Valappil SP, Roy I, Boccaccini AR (2006). Polyhydroxyalkanoate (PHA)/inorganic phase composites for tissue engineering applications. Biomacromolecules 7: 2249– 2258.
Novikov LN, Novikova LN, Mosahebi A, Wiberg M, Terenghi G, Kellerth JO (2002). A novel biodegradable implant for neuronal rescue and regeneration after spinal cord injury. Biomaterials 23: 3369–3376.
Novikova LN, Pettersson J, Brohlin M, Wiberg M, Novikov LN (2008). Biodegradable poly-beta-hydroxybutyrate scaffold seeded with Schwann cells to promote spinal cord repair. Biomaterials 29: 1198–1206.
Palmquist A, Omar OM, Esposito M, Lausmaa J, Thomsen P (2010). Titanium oral implants: surface characteristics, interface biology and clinical outcome. J Roy Soc Interface 7 (Suppl 5): S515–S527.
Rentsch C, Rentsch B, Breier A, Hofmann A, Manthey S, Scharnweber D, Biewener A, Zwipp H (2010). Evaluation of the osteogenic potential and vascularization of 3D poly(3)hydroxybutyrate scaffolds subcutaneously implanted in nude rats. J Biomed Mater Res A 92: 185–195.
Saito T, Tomita K, Juni K, Ooba K (1991). In vivo and in vitro degradation of poly(hydroxybutyrate) in rat. Biomaterials 12: 309–312.
Sangsanoh P, Waleetorncheepsawat S, Suwantong O, Wutticharoenmongkol P, Weeranantanapan O, Chuenjitbuntaworn B, Cheepsunthorn P, Pavasant P, Supaphol P (2007). In vitro biocompatibility of schwann cells on surfaces of biocompatible polymeric electrospun fibrous and solution-cast film scaffolds. Biomacromolecules 8: 1587–1594.
Shishatskaya EI, Volova TG (2004). A comparative investigation of biodegradable polyhydroxyalkanoate films as matrices for in vitro cell cultures. J Mater Sci-Mater M 15: 915–923.
Suwantong O, Waleetorncheepsawat S, Sanchavanakit N, Pavasant P, Cheepsunthorn P, Bunaprasert T, Supaphol P (2007). In vitro biocompatibility of electrospun poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) fiber mats. Int J Biol Macromol 40: 217–223.
Takahashi Y, Tabata Y (2003). Homogeneous seeding of mesenchymal stem cells into nonwoven fabric for tissue engineering. Tissue Eng 9: 931–938.
Wang YW, Wu Q, Chen GQ (2004). Attachment, proliferation and differentiation of osteoblasts on random biopolyester poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) scaffolds. Biomaterials 25: 669–675.
Wei X, Hu YJ, Xie WP, Lin RL, Chen GQ (2009). Influence of poly(3-hydroxybutyrate-co-4-hydroxybutyrate-co-3- hydroxyhexanoate) on growth and osteogenic differentiation of human bone marrow-derived mesenchymal stem cells. J Biomed Mater Res A 90: 894–905.
Wollenweber M, Domaschke H, Hanke T, Boxberger S, Schmack G, Gliesche K, Scharnweber D, Worch H (2006). Mimicked bioartificial matrix containing chondroitin sulphate on a textile scaffold of poly(3-hydroxybutyrate) alters the differentiation of adult human mesenchymal stem cells. Tissue Eng 12: 345–359.
Wu Q, Wang Y, Chen GQ (2009). Medical application of microbial biopolyesters polyhydroxyalkanoates. Artif Cell Blood Sub 37: 1–12.
Xu F, Shi J, Yu B, Ni W, Wu X, Gu Z (2010). Chemokines mediate mesenchymal stem cell migration toward gliomas in vitro. Oncol Rep 23: 1561–1567.
Yang X, Zhao K, Chen GQ (2002). Effect of surface treatment on the biocompatibility of microbial polyhydroxyalkanoates. Biomaterials 23: 1391–1397.
Yasin M, Tighe BJ (1992). Polymers for biodegradable medical devices. VIII. Hydroxybutyrate-hydroxyvalerate copolymers: physical and degradative properties of blends with polycaprolactone. Biomaterials 13: 9–16.
Young RC, Wiberg M, Terenghi G (2002). Poly-3- hydroxybutyrate (PHB): a resorbable conduit for long-gap repair in peripheral nerves. Brit J Plast Surg 55: 235–240.
Yu B, Chen P, Sun Y, Lee Y, Young T (2010). Effects of the surface characteristics of polyhydroxyalkanoates on the metabolic activities and morphology of human mesenchymal stem cells. J Biomat Sci-Polym E 21: 17–36.
Yu Q, Xu S, Zhang H, Gu L, Xu Y, Ko F (2013). Structure-property relationship of regenerated spider silk protein nano/ microfibrous scaffold fabricated by electrospinning. J Biomed Mater Res A 102: 3828–3837