Induction of Angiogenesis by Matrigel Coating of VEGF-Loaded PEG/PCL-Based Hydrogel Scaffolds for hBMSC Transplantation

다국어 초록 (Multilingual Abstract)

hBMSCs are multipotent cells that are useful for tissue regeneration to treat degenerative diseases and others for their differentiation ability into chondrocytes, osteoblasts, adipocytes, hepatocytes and neuronal cells. In this study, biodegradable elastic hydrogels consisting of hydrophilic poly(ethylene glycol) (PEG) and hydrophobic poly(- caprolactone) (PCL) scaffolds were evaluated for tissue engineering because of its biocompatibility and the ability to control the release of bioactive peptides. The primary cultured cells from human bone marrow are confirmed as hBMSC by immunohistochemical analysis. Mesenchymal stem cell markers (collagen type I, fibronectin, CD54, integrin1 , and Hu protein) were shown to be positive, while hematopoietic stem cell markers (CD14 and CD45) were shown to be negative. Three different hydrogel scaffolds with different block compositions (PEG:PCL=6:14 and 14:6 by weight) were fabricated using the salt leaching method.
The hBMSCs were expanded, seeded on the scaffolds, and cultured up to 8 days under static conditions in Iscove's Modified Dulbecco's Media (IMDM). The growth of MSCs cultured on the hydrogel with PEG/PCL= 6/14 was faster than that of the others. In addition, the morphology of MSCs seemed to be normal and no cytotoxicity was found.
The coating of the vascular endothelial growth factor (VEGF) containing scaffold with Matrigel slowed down the release of VEGF in vitro and promoted the angiogene-sis when transplanted into BALB/c nude mice. These results suggest that hBMSCs can be supported by a biode gradable hydrogel scaffold for effective cell growth, and enhance the angiogenesis by Matrigel coating.

참고문헌 (Reference)

1 Tabata, Y., "Vascularization effect of basic fibroblast growth factor released from gelatin hydrogels with different biodegradabilities" 20 : 2169-2175, 1999

2 Mehdizadeh, H., "Three-dimensional modeling of angiogenesis in porous biomaterial scaffolds" 34 : 2875-2887, 2013

3 Kanczler, J. M., "The effect of mesenchymal populations and vascular endothelial growth factor delivered from biodegradable polymer scaffolds on bone formation" 29 : 1892-1900, 2008

4 Su Jin Im, "Synthesis and Characterization of Biodegradable Elastic Hydrogels Based on Poly(ethylene glycol) and Poly(ε-caprolactone) Blocks" 한국고분자학회 15 (15): 363-369, 2007

5 Kanczler, J. M., "Supercritical carbon dioxide generated vascular endothelial growth factor encapsulated poly(DL-lactic acid)scaffolds induce angiogenesis in vitro" 352 : 135-141, 2007

6 Schantz, J. T., "Repair of calvarial defects with customised tissueengineered bone grafts II. Evaluation of cellular efficiency and efficacy in vivo" 9 (9): 127-139, 2003

7 Khare, A. R., "Release behavior of bioactive agents from pH-sensitive hydrogels" 4 : 275-289, 1993

8 da Silva, G. R., "Polyurethanes as supports for human retinal pigment epithelium cell growth" 34 : 198-209, 2011

9 Costa-Pinto, A. R., "Osteogenic differentiation of human bone marrow mesenchymal stem cells seeded on melt based chitosan scaffolds for bone tissue engineering applications" 10 : 2067-2073, 2009

10 GREEN DAVID WILLIAM, "Osteogenic Potency of Nacre on Human Mesenchymal Stem Cells" 한국분자세포생물학회 38 (38): 267-272, 2015

11 Lee, H. Y., "Novel porous scaffolds of poly(lactic acid)produced by phaseseparation using room temperature ionic liquid and the assessments of biocompatibility" 23 : 1271-1279, 2012

12 Pittenger, M. F., "Multilineage potential of adult human mesenchymal stem cells" 284 : 143-147, 1999

13 Zanatta, G., "Mesenchymal stem cell adherence on poly(D, L-lactide-co-glycolide)nanofibers scaffold is integrin-beta 1 receptor dependent" 8 : 211-218, 2012

14 Kleinman, H. K., "Matrigel : basement membrane matrix with biological activity" 15 : 378-386, 2005

15 De Kok, I. J., "Investigation of allogeneic mesenchymal stem cell-based alveolar bone formation : preliminary findings" 14 : 481-489, 2003

16 Laschke, M. W., "Incorporation of growth factor containing Matrigel promotes vascularization of porous PLGA scaffolds" 85 : 397-407, 2008

17 Mankani, M. H., "In vivo bone formation by human bone marrow stromal cells : effect of carrier particle size and shape" 72 : 96-107, 2001

18 Park, J. S., "In vitro and in vivo test of PEG/PCL-based hydrogel scaffold for cell delivery application" 124 : 51-59, 2007

19 Zeiser, R., "Immunopathogenesis of acute graft-versus-host disease : implications for novel preventive and therapeutic strategies" 83 : 551-565, 2004

20 Lee, K. Y., "Hydrogels for tissue engineering" 101 : 1869-1879, 2001

21 Ho, Y. C., "Heparinfunctionalized chitosan-alginate scaffolds for controlled release of growth factor" 376 : 69-75, 2009

22 Bruder, S. P., "Growth kinetics, self-renewal, and the osteogenic potential of purified human mesenchymal stem cells during extensive subcultivation and following cryopreservation" 64 : 278-294, 1997

23 Yang, F., "Genetic engineering of human stem cells for enhanced angiogenesis using biodegradable polymeric nanoparticles" 107 : 3317-3322, 2010

24 Yang, X. B., "Evaluation of human bone marrow stromal cell growth on biodegradable polymer/bioglass composites" 342 : 1098-1107, 2006

25 Li, W. J., "Evaluation of articular cartilage repair using biodegradable nanofibrous scaffolds in a swine model : a pilot study" 3 : 1-10, 2009

26 Perets, A., "Enhancing the vascularization of threedimensional porous alginate scaffolds by incorporating controlled release basic fibroblast growth factor microspheres" 65 : 489-497, 2003

27 Peters, M. C., "Engineering vascular networks in porous polymer matrices" 60 : 668-678, 2002

28 Tsuchida, H., "Engineered allogeneic mesenchymal stem cells repair femoral segmental defect in rats" 21 : 44-53, 2003

29 Lin, J., "Effects of surface-modified scaffolds on the growth and differentiation of mouse adipose-derived stromal cells" 1 : 211-217, 2007

30 Heo, J. Y., "Downregulation of APE1/Ref-1 is involved in the senescence of mesenchymal stem cells" 27 : 1455-1462, 2009

31 Phinney, D. G., "Concise review : mesenchymal stem/multipotent stromal cells : the state of transdifferentiation and modes of tissue repair--current views" 25 : 2896-2902, 2007

32 Chen, D. C., "Comparison of polyester scaffolds for bioengineered intestinal mucosa" 184 : 154-165, 2006

33 Park, J. S., "Chondrogenic differentiation of mesenchymal stem cells embedded in a scaffold by long-term release of TGF-beta 3complexed with chondroitin sulfate" 92 : 806-816, 2010

34 Kim, H. J., "Chondrogenesis using mesenchymal stem cells and PCL scaffolds" 92 : 659-666, 2010

35 Tanaka, T., "Bone augmentation by bone marrow mesenchymal stem cells cultured in threedimensional biodegradable polymer scaffolds" 91 : 428-435, 2009

36 Duailibi, M. T., "Bioengineered teeth from cultured rat tooth bud cells" 83 : 523-528, 2004

37 Schmidmaier, G., "Biodegradable poly(D, L-lactide)coating of implants for continuous release of growth factors" 58 : 449-455, 2001

38 Hennink, W. E., "Biodegradable dextran hydrogels crosslinked by stereocomplex formation for the controlled release of pharmaceutical proteins" 277 : 99-104, 2004

39 Papo, N., "Antagonistic VEGF variants engineered to simultaneously bind to and inhibit VEGFR2 and alphavbeta3 integrin" 108 : 14067-14072, 2011

40 Yang, X., "Angiogenesis defects and mesenchymal apoptosis in mice lacking SMAD5" 126 : 1571-1580, 1999

41 Deng, C., "A collagenchitosan hydrogel for endothelial differentiation and angiogenesis" 16 : 3099-3109, 2010

42 Zhijian Zhang, "A Fibrin Matrix Promotes the Differentiation of EMSCs Isolated from Nasal Respiratory Mucosa to Myelinating Phenotypical Schwann-Like Cells" 한국분자세포생물학회 38 (38): 221-228, 2015