Evaluation of Stem Cells’ Growth on Electrospun Polycaprolactone (PCL) Scaffolds Used for Soft Tissue Applications

Published: 2021-07-07 00:09:43
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IntroductionStem cells can be defined as versatile, unspecialized cells that have the potential either to divide to make more stem cells or to differentiate into one or more cell type, usually in response to some kind of signal (Evans, 2006).Tissue engineering can be defined as using a scaffold with the appropriate architecture to guide the repair and restoration of function to damaged tissues or organs (Yao et al., 2011).Developing scaffolds that mimic the architecture of normal tissue is one of the major challenges in the field of tissue engineering (Shoichet, 2009).Nowadays, there are several techniques available for the synthesis that matches such challenges (Johnson et al., 2009)Electrospinning is the most widely studied technique and has also demonstrated the most promising results in terms of tissue engineering applications (Athira et al., 2014).ArticleEvaluation of Stem Cells’ Growth on Electrospun Polycaprolactone (PCL) Scaffolds Used for Soft Tissue Applications (Akram et al., 2017).GoalsThis study focuses on the preparation of a synthetic tissue from a biodegradable polymer that can be used for soft tissue applications using electrospinning technique.Approach / Procedure / MethodsExperimental technique has been adopted in this paper.ResultsScanning electron microscope image for the processed (Fig. 2(a)) 10 % W/V PCL scaffold, showed that the low polymer concentration result in small fiber diameter since the average fiber diameter was about 259 nm (Fig. 2(b)) and this will provide the large surface area favorable for cell adhesion.There is a decrease in the contact angle with time (Table 1); therefore, the scaffolds were soaked in media for two hours after soaking in serum and before adding stem cells in order to be more hydrophilic and to improve the cells attachment.SEM analysis revealed low growth rates for stem cells, and this may attributed to the beadon- string structure for this scaffold which will decrease the surface area allowed for cell adhesion, because increasing the fiber diameter will decrease the surface area and this in turn will lower cell adhesion (Li et al., 2005).Histological analysis revealed that the presence of the beaded structure prevented the spread of cells inside the scaffold, despite the good average pore size for this scaffold which was equal to 6400 nm.The formation of confluent monolayer of stem cells was noticed with the scaffold 15 % W/V PCL soaked in serum.The staining with toluidine blue for 15% PCL scaffold which was not soaked in serum also revealed the suitability of this scaffold structure for stem cells spread inside the scaffold.ConclusionThe results of cell growth analysis by SEM for the prepared scaffolds which are (10, 15, and 20) % W/V PCL, showed that the best scaffold in supporting stem cells adhesion and proliferation was 15 % W/V PCL, because it revealed high growth rates of stem cells even when it wasn’t soaked in serum.However, soaking the scaffolds in 100% fetal bovine serum before adding the media with cells, will improve the hydrophilicity of the scaffold surface which will improve in turn the attachment of cells on the scaffold and accelerated the formation of confluent monolayer.On the other side, the results of histological analysis for these scaffolds revealed the suitability of uniform fibrous structure that is beads free, with pore size larger than 5000 nm for cell spread inside the scaffold.This was noticed in both 15 and 20% PCL scaffolds, which had uniform fibrous structures without beads, and average pore size larger than 5000 nm.Therefore, both 15 and 20% PCL scaffolds supported the spread of stem cells inside the scaffold to enhance the 3D growth and the formation of a regenerative tissue.ReferencesAthira KS, Sanpui P, Chatterjee K. Fabrication of Poly (Caprolactone) Nanofibers by Electrospinning. Journal of Polymer and Biopolymer Physics Chemistry; 2014. 2:62-6.Chen- Ming Hsu, Electrospinning of poly (ε- Caprolactone), Master thesis, Faculty of Worcester poly technic Institute; 2003.Croisier F, Duwez AS, Jérôme C, Léonard AF, Van Der Werf KO, Dijkstra PJ, Bennink ML. Mechanical testing of electrospun PCL fibers. Acta biomaterialia; 2012. 8: 218-24da Silva MA, Crawford A, Mundy J, Martins A, Araújo JV, Hatton PV, Reis RL, Neves NM. Evaluation of extracellular matrix formation in polycaprolactone and starch-compounded polycaprolactone nanofiber meshes when seeded with bovine articular chondrocytes. Tissue Engineering Part A; 2008 .15: 377-85.Evans ND, Gentleman E, Polak JM. Scaffolds for stem cells. Materials Today; 2006. 9: p.26-33Fromstein JD, Zandstra PW, Alperin C, Rockwood D, Rabolt JF, Woodhouse KA. Seeding bioreactor-produced embryonic stem cell-derived cardiomyocytes on different porous, degradable, polyurethane scaffolds reveals the effect of scaffold architecture on cell morphology. Tissue Engineering Part A; 2008. 14: 369-78.Gluck Jessica Marie, Electrospun nanofibrous poly (ε- caprolactone) (PCL) scaffold for liver tissue engineering, MSc. Thesis, Faculty of North Carolina State University; 2007.Johnson J, Niehaus A, Nichols S, Lee D, Koepsel J, Anderson D, Lannutti J. Electrospun PCL in vitro: a microstructural basis for mechanical property changes. Journal of Biomaterials Science, Polymer Edition; 2009 .20: p. 467-81.Li M, Mondrinos MJ, Gandhi MR, Ko FK, Weiss AS, Lelkes PI. Electrospun protein fibers as matrices for tissue engineering. Biomaterials; 2005. 26: p. 5999-6008.Milleret V, Hefti T, Hall H, Vogel V, Eberli D. Influence of the fiber diameter and surface roughness of electrospun vascular grafts on blood activation. Acta biomaterialia; 2012. 8: 4349-56.Oliveira JT, Crawford A, Mundy JM, Moreira AR, Gomes ME, Hatton PV, Reis RL. A cartilage tissue engineering approach combining starch-polycaprolactone fibre mesh scaffolds with bovine articular chondrocytes. Journal of Materials Science: Materials in Medicine; 2007 .18: p. 295-302.Shoichet MS. Polymer scaffolds for biomaterials applications. Macromolecules; 2009 .43: p. 581-91.Yao J, Tao SL, Young MJ. Synthetic polymer scaffolds for stem cell transplantation in retinal tissue engineering. Polymers; 2011. 3: p.899- 914.

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