3D nanostructured microcarriers for cell therapy in regenerative medicine

Della Porta, Giovanna (2013) 3D nanostructured microcarriers for cell therapy in regenerative medicine, [Dissertation thesis], Alma Mater Studiorum Università di Bologna. Dottorato di ricerca in Bioingegneria, 25 Ciclo. DOI 10.6092/unibo/amsdottorato/5327.
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Abstract

Supercritical Emulsion Extraction technology (SEE-C) was proposed for the production of poly-lactic-co-glycolic acid microcarriers. SEE-C operating parameters as pressure, temperature and flow rate ratios were analyzed and the process performance was optimized in terms of size distribution and encapsulation efficiency. Microdevices loaded with bovine serum insulin were produced with different sizes (2 and 3 µm) or insulin charges (3 and 6 mg/g) and with an encapsulation efficiency of 60%. The microcarriers were characterized in terms of insulin release profile in two different media (PBS and DMEM) and the diffusion and degradation constants were also estimated by using a mathematical model. PLGA microdevices were also used in a cultivation of embryonic ventricular myoblasts (cell line H9c2 obtained from rat) in a FBS serum free medium to monitor cell viability and growth in dependence of insulin released. Good cell viability and growth were observed on 3 µm microdevices loaded with 3 mg/g of insulin. PLGA microspheres loaded with growth factors (GFs) were charged into alginate scaffold with human Mesenchimal Steam Cells (hMSC) for bone tissue engineering with the aim of monitoring the effect of the local release of these signals on cells differentiation. These “living” 3D scaffolds were incubated in a direct perfusion tubular bioreactor to enhance nutrient transport and exposing the cells to a given shear stress. Different GFs such as, h-VEGF, h-BMP2 and a mix of two (ratio 1:1) were loaded and alginate beads were recovered from dynamic (tubular perfusion system bioreactor) and static culture at different time points (1st, 7th, 21st days) for the analytical assays such as, live/dead; alkaline phosphatase; osteocalcin; osteopontin and Van Kossa Immunoassay. The immunoassay confirmed always a better cells differentiation in the bioreactor with respect to the static culture and revealed a great influence of the BMP-2 released in the scaffold on cell differentiation.

Abstract
Tipologia del documento
Tesi di dottorato
Autore
Della Porta, Giovanna
Supervisore
Co-supervisore
Dottorato di ricerca
Scuola di dottorato
Scienze e ingegneria dell'informazione
Ciclo
25
Coordinatore
Settore disciplinare
Settore concorsuale
Parole chiave
Tissue engineering, scaffold, biopolymer, osteoblast
URN:NBN
DOI
10.6092/unibo/amsdottorato/5327
Data di discussione
12 Aprile 2013
URI

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