A reproducible, high throughput method for fabricating fibrin gels

BMC Research Notes

J Kent Leach¹  Kaitlin C Murphy¹ 

[1] Department of Biomedical Engineering, University of California, Davis, 451 Health Sciences Drive, Davis, CA, 95616, USA

关键词: Bone;    Mesenchymal stem cells;    Polydimethylsiloxane;    Hydrogel;    Fibrin;   

Others  :  1165971
DOI  :  10.1186/1756-0500-5-423

 received in 2012-06-21, accepted in 2012-07-31,  发布年份 2012

【 摘 要 】

Background

Fibrin gels are a promising biomaterial for tissue engineering. However, current fabrication methods are time intensive with inherent variation. There is a pressing need to develop new and consistent approaches for producing fibrin-based hydrogels for examination.

Findings

We developed a high throughput method for creating fibrin gels using molds fabricated from polydimethylsiloxane (PDMS). Fibrin gels were produced by adding solutions of fibrinogen and thrombin to cylindrical defects in a PDMS sheet. Undisturbed gels were collected by removing the sheet, and fibrin gels were characterized. The characteristics of resulting gels were compared to published data by measuring compressive stiffness and osteogenic response of entrapped human mesenchymal stem cells (MSCs). Gels exhibited compressive moduli nearly identical to our previously reported fabrication method. Trends in alkaline phosphatase activity, an early marker of osteogenic differentiation in MSCs, were also consistent with previous data.

Conclusions

These findings demonstrate a streamlined approach to fibrin gel production that drastically reduces the time required to make fibrin gels, while also reducing variability between gel batches. This fabrication technique provides a valuable tool for generating large numbers of gels in a cost-effective manner.

【 授权许可】

   
2012 Murphy and Leach; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Janmey P, Winer JP, Weisel JW: Fibrin gels and their clinical and bioengineering applications. J R Soc Interface 2009, 6:1-10.
• [2]Shaikh FM, Callanan A, Kavanagh EG, Burke PE, Grace PA, McGloughlin TM: Fibrin: a natural biodegradable scaffold in vascular tissue engineering. Cells Tissues Organs 2008, 188:333-346.
• [3]Barsotti M, et al.: Fibrin acts as biomimetic niche inducing both differentiation and stem cell marker expression of early human endothelial progenitor cells. Cell Prolif 2011, 44:33-48.
• [4]Hubbell JA: Biomaterials in tissue engineering. Nat Biotechnol 1995, 13:565-576.
• [5]Grassl ED, Oegema TR, Tranquillo RT: Fibrin as an alternative biopolymer to type-1 collagen for the fabrication of a media equivalent. J Biomed Mater Res 2002, 60:607-612.
• [6]Bensaïd W, Triffitt JT, Blanchat C, Oudina K, Sedel L, Petite H: A biodegradable fibrin scaffold for mesenchymal stem cell transplantation. Biomaterials 2003, 24:2497-2502.
• [7]Siebenlist KR, Mosesson MW: Progressive cross-linking of fibrin gamma chains increases resistance to fibrinolysis. J Biol Chem 1994, 269:28414-28419.
• [8]Blombäck B, Bark N: Fibrinopeptides and fibrin gel structure. Biophys Chem 2004, 112:147-151.
• [9]Carr ME, Gabriel DA, McDonagh J: Influence of Ca2+ on the structure of reptilase-derived and thrombin-derived fibrin gels. Biochem J 1986, 239:513-516.
• [10]Okada M, Blomback B: Calcium and fibrin gel structure. Thromb Res 1983, 29:269-280.
• [11]Davis HE, Miller SL, Case EM, Leach JK: Supplementation of fibrin gels with sodium chloride enhances physical properties and ensuing osteogenic response. Acta Biomater 2011, 7:691-699.
• [12]Zhao HG, Ma L, Zhou J, Mao ZW, Gao CY, Shen JC: Fabrication and physical and biological properties of fibrin gel derived from human plasma. Biomed Mater 2008, 3:15001-15010.
• [13]Marois Y, Be MC: Studies of primary reference materials low-density polyethylene and polydimethylsiloxane : A review. J Biomed Mater Res 2001, 58:467-477.
• [14]Regehr KJ, et al.: Biological implications of polydimethylsiloxane-based microfluidic cell culture. Lab Chip 2009, 9:2132-2139.