Abstract
Polymeric microspheres are versatile tissue engineering scaffold, suitable for simultaneous drug delivery and surface modification. They are small units that can be individually tailored for different cell types and purposes, then assembled with various compositions and shapes. In addition, they provide a 3D environment for cell-cell and cell-material interactions. Poly(3-hydroxyburate-co-3-hydroxyvalerate) (PHBV, 8% PHV content), a biodegradable and biocompatible microbial polyester, was used to fabricate microspheres using an oil-in-water emulsion solvent evaporation technique. As PHBV is biodegradable, the eventual construct will consist of only natural tissues, avoiding chronic complications. The versatility of microspheres is useful in neural therapies which require several cell types and various biomolecular signals to overcome inflammation, glial scarring, and inhibitory signals, as well as to promote repair and regeneration. Due to cell source limitations, neuronal expansion might have to be carried out on the microspheres to yield enough neurons for therapies. Also, a key requirement for engineered neural tissue would be its functional integration into host tissue. Thus, the suitability of PHBV microspheres as neural tissue engineering scaffolds was investigated using neuro2a neuroblastoma cells as a model for proliferating neurons and primary mouse fetal cortical neurons as a model for functioning neurons. Scanning electron microscope (SEM) imaging, and confocal laser scanning microscopy were used to observe morphologies of cells cultured on PHBV microspheres. Also, DNA Quantification and cell viability assay (MTT assay) were carried out. Neuro2a cultures on PHBV microspheres showed a growth trend comparable to that in 2D culture plate controls, in both assays. Also, through immunoflourescence staining for beta-III tubulin, primary neurons were found to extend neurites on PHBV microspheres. The results showed that PHBV microspheres are suitable scaffolds for neural tissue engineering, as they support the neural cells’ adhesion, growth, proliferation and differentiation.
Keywords
- Neural Tissue Engineering
- Polymeric microspheres
- Poly(3-hydroxyburate-co-3-hydroxyvalerate)
This is a preview of subscription content, access via your institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsPreview
Unable to display preview. Download preview PDF.
References
Ofira E, Tamir BH. The changing face of neural stem cell therapy in neurologic diseases. Arch neurol 2008; 65(4): 452–456.
Guzman R, Choi R, Gera A, Angeles ADL, Andres RH, Steinberg GK. Intravascular cell replacement therapy for stroke. Neurosurg Focus 2008; 24(3&4): E14.
Eftekharpour E, Karimi-Abdolrezaee S, Fehlings MG. Current status of experimental cell replacement approaches to spinal cord injury. Neurosurg Focus 2008; 24(3&4): E18.
Chen CP, Kiel ME, Sadowski D, McKinnon RD. From stem cells to oligodendrocytes: Prospects for Brain Therapy. Stem Cell Rev 2007; 3: 280–288.
O’Donnell PB, McGinit JW. Preparation of microspheres by the solvent evaporation technique. Adv Drug Deliv Rev 1997; 28: 25–42.
Xu XJ, Sy JC, Prasad SV. Towards developing surface eroding PHAs. Biomaterials 2006; 27(15): 3012–3030.
Chen J, Davis SS. The release of diazepam from poly(hydroxybutyrate-hydroxyvalerate) microspheres. J Microencapsul 2002: 19(2): 191–201.
Van Tomme SR, Van Steenbergen MJ, De Smedt SC, Van Nostrum CF, Hennink WE. Self-gelling hydrogels based on oppositely charged dextran microspheres. Biomaterials 2005; 26(14): 2129–2135.
Payne RG, Yasko AW, Yaszemski MJ, Mikos AG. Temporary encapsulation of rat marrow osteoblasts in gelatin microspheres for bone tissue engineering. Materials Research Society Symposium — Proceedings (2001).
Wong MS, Causey TB, Mantzaris N, Bennett GN, San KY. Engineering poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer composition in E. Coli. Biotechnol Bioeng 2008; 99(4): 919–928.
Squio CR, Marangoni C, De Vecchi CS, Aragao GMF. Phosphate feeding strategy during production phase improves poly(3-hydroxybutyrate-co-3-hydroxyvalerate) storage by Ralstonia eutropha. Appl Microbiol Biotechnol 2003; 61: 257–260.
Hankermeyer CR, Tjeerdema Rs. Polyhydroxyburate: plastic made and degraded by microorganisms. Rev Environ Contam Toxicol 1999; 159: 1–24.
Gürsel I, Korkusuz F, Türesin F, Alaeddinoglu N.G, Hasýrcý V. In vivo application of biodegradable controlled antibiotic release systems for the treatment of implant-related osteomyelitis. Biomaterials 2001; 22: 73–80.
Willerth SM, Sakiyama-Elbert SE. Approaches to neural tissue engineering using scaffolds for drug delivery. Adv Drug Deliv Rev 2007; 59: 325–338.
Kallos MS, Sen A, Behie LA. Large scale expansion of mammalian neural stem cells: a review. Med Biol Eng Comput 2003; 41: 271–282.
Schmidt CE, Leach JB. Neural tissue engineering: strategies for repair and regeneration. Annu Rev Biomed Eng 2003; 5: 293–347.
Yang YY, Chia HH, Chung TS. Effect of preparation temperature on the characteristics and release profile of PLGA microspheres containing protein fabricated by double-emulsion solvent extraction/evapouration method. J Control Release 2000; 69: 81–96.
Ciapetti G, Cenni E, Pratelli L, Pizzoferrato A. In vitro evaluation of cell/biomaterial interaction by MTT assay. Biomaterials 1993: 14(5): 359–364.
Rago R, Mitchen J, Wilding G. DNA fluorometric assay in 96-well tissue culture plates using Hoechst 33258 after cell lysis by freezing in distilled water. Anal Biochem 1990; 191: 31–34.
Moskowitz PF, Oblinger MM. Sensory neurons selectively upregulate synthesis and transport of the beta-III-tubulin protein during axonal regeneration. J neurosci 1995; 15(2): 1545–1555.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 International Federation of Medical and Biological Engineering
About this paper
Cite this paper
Chen, W.H., Tang, B.L., Tong, Y.W. (2009). PHBV Microspheres as Tissue Engineering Scaffold for Neurons. In: Lim, C.T., Goh, J.C.H. (eds) 13th International Conference on Biomedical Engineering. IFMBE Proceedings, vol 23. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-540-92841-6_296
Download citation
DOI: https://doi.org/10.1007/978-3-540-92841-6_296
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-540-92840-9
Online ISBN: 978-3-540-92841-6
eBook Packages: EngineeringEngineering (R0)