@article {19273, title = {Chondrogenesis-inductive nanofibrous substrate~comprising autologous TGF-b3 and IGF-I from platelet lysates}, journal = {GENE2SKIN~Conference \& TERM STEM 2016}, year = {2016}, month = {2016-10-24 00:00:00}, address = {Guimar{\~a}es,~Portugal}, abstract = {

Articular cartilage is a connective tissue with low self-regeneration potential due to its avascular nature and lack of progenitor cells [1]. Therefore, cartilage regeneration is far to be achieved and then it is an important candidate for tissue engineering and regenerative medicine (TERM) strategies. TGF-b3 and IGF-I have been identified as important proteins on the regulation of cartilage development and on the homeostasis of mature articular cartilage [2, 3]. Therefore, the synergetic effect of the growth factors (GFs) in a biomaterial substrate is hypothesized to lead to the successful chondrogenic differentiation of mesenchymal stem cells (MSCs). The leading goal of this study is to develop a multi-functionalized electrospun nanofibrous mesh (NFM) with chondrogenic potential, through the binding of autologous TGF-b3 and IGF-I. For that, defined antibodies were immobilized at the surface of NFMs, capable to specifically select only the GFs of interest from a complex biological fluid (i.e. platelet lysate - PL). The maximum immobilization capacity of each antibody was 4 mg/mL for both antibodies. The anti-TGF-b3 and anti-IGF-I antibodies were also immobilized over the same structure in a mixed fashion at 1:10 proportion (0.4 mg/mL anti-TGF-b3 : 3.6 mg/mL anti-IGF-I), which corresponds to the proportion used in the chondrogenic differentiation medium. The bioactivity of immobilized antibodies was tested by using recombinant proteins, achieving a binding efficiency of 36\% for TGF-b3 and 44\% for IGF-I. Similar results were observed for GFs captured from PL by NFMs functionalized with single or mixed antibodies. The chondrogenic potential of this bi-functionalized biomaterial system (Single or Mixed, with TGF-b3 and IGF-I captured from PL or recombinant proteins) was further assessed by culturing human bone marrow-derived MSCs during 28 days without further induction. Bare NFMs cultured with hBMSCs under standard chondrogenic differentiation medium supplemented with TGF-b3 and IGF-I were used as positive control. Biological results indicate that the bio-functional systems are more effective when compared to control condition, as confirmed either by histology and by real time-PCR quantification. Furthermore, and interestingly, the results show a stronger chondrogenic induction by the GFs from PL as compared to the recombinant GFs.

[1] Correa, D. and S.A. Lietman, Semin Cell Dev Biol, 2016;

[2] Petrou, M., et al., Regen Med, 2013;

[3] Longobardi, L., et al., J Bone Miner Res, 2006.

}, keywords = {chondrogenic differentiation, growth factors, Nanofibrous meshes, Platelet lysate}, author = {Casanova, M. R. and Alves da Silva, ML and Costa-Pinto, A. R. and Reis, R. L. and Martins, A. and Neves, N. M.} }