http://informahealthcare.com/plt ISSN: 0953-7104 (print), 1369-1635 (electronic) Platelets, 2014; 25(7): 556–557 ! 2014 Informa UK Ltd. DOI: 10.3109/09537104.2013.836748
LETTER
Platelet and plasma bioactive scaffolds for stem cell differentiation: What are we missing? Andrei Moroz1,2, Se´rgio L. Felisbino2, & Elenice Deffune1 1
Cell Engineering Lab, Blood Transfusion Center, Botucatu Medical School, Univ Estadual Paulista, UNESP, Botucatu, SP, Brazil and 2Extracellular Matrix Lab, Department of Morphology, Botucatu Biosciences Institute, Univ Estadual Paulista, UNESP, Botucatu, SP, Brazil
To the editor, Tissue engineering has evolved rapidly over the past 10 years and hold great promise for the treatment of several health injuries, especially due to the accumulated knowledge in stem cell biology, cultivation techniques and differentiation capabilities [1]. Many tissues to be generated in vitro require three dimensional (3D) scaffolds for the successful differentiation of stem cells into the target/desired cell phenotype, such as cartilage tissue and others [1, 2]. Due to this escalating demand, scaffolding technology for cell culture/therapy now allows for more efficient approaches, notably the employment of bioactive materials as their major constituents [2]. One recently published report at this journal demonstrated the feasibility of using platelets as the main component of a bioactive 3D constructed scaffold for mesenchymal stem cell (MSC) cultivation [2]. This scaffold has been found to be suitable for the differentiation of the referred cells in chondrocytes, as demonstrated using extracellular matrix characterization [2] and molecular biology techniques [3]. Moreover, its main component is easy to access, especially in blood transfusion centers. Although this scaffold has been systematically considered to be bioactive largely due to the growth factors, such as PDGF, VEGF, FGF, TGF-b, and others, present at platelets’ granules [3], to date, no one has ever considered a role for plasma fibronectin, as a bioactive agent, which could deliver extra differentiation cues for stem cells cultivated inside these scaffolds. Interestingly, accumulated evidence shows that fibronectin, an abundant protein present at blood plasma, bounded to platelets, and extracellular matrix (ECM), is able to actively modify the biology behavior of both stem [4] and tumoral cells [5]. In this sense, it has been reported that traits such as the activity of matrix metalloproteinases (MMPs), important endopeptidases related to ECM remodeling, which could not be detected at the conditioned medium of prostate normal and tumoral human cells, rapidly increases after short time exposure to fibronectin [5]. Given that the 3D scaffold structure needs to be reabsorbed, as the EMC proteins are produced by the seeded cells, this stimulus on MMP activity could be beneficial during the remodeling of the 3D structure. Moreover, stem cells cultivated at bone differentiation Correspondence: A. Moroz, Univ Estadual Paulista, UNESP, Botucatu Medical School, Blood Transfusion Center, Cell Engineering Lab, District of Rubia˜o Ju´nior S/N, 18618-970, Botucatu, SP, Brazil. Tel: +551438116041 (r 235). E-mail: [email protected]
Keywords Bioactive scaffolds, extracellular matrix, fibronectin, platelet based scaffolds, platelet-rich plasma, stem cells History Received 22 July 2013 Revised 15 August 2013 Accepted 18 August 2013 Published online 31 October 2013
medium exhibited enhanced alkaline phosphatase activity and calcium deposition after exposure to fibronectin coated to culture plates [4], which backs up a possible differentiation role of fibronectin in platelet-derived scaffolds, as a synergistic inductor effect to the stimulus provided from the aforementioned growth factors. Also very relevant, other bioactive scaffolds such as those derived from fibrin sealants are commonly used for stem cell differentiation protocols [6]. For a detailed review on these scaffolds and their applications for regenerative medicine, readers are advised to seek a recently published report [7]. These products, which are commercially available [8, 9] or obtained with an automated device for the production of fibrin sealant [9], have been characterized (nevertheless of the method of obtention) as being enriched in fibronectin content, as observed through the use of specific antibodies and sodium dodecyl sulphate– polyacrylamide gel electrophoresis [9]. Therefore, protocols that employ these scaffolds should also take into consideration the possible influence of fibronectin, on the stimulus provided to the differentiation of the seeded stem cells and their MMPs expression/activity. However, this protein could also exert a proliferative induction of stem cells cultivated in platelet constructed scaffolds, as there are reports of higher proliferation of CD34þ cells when fibronectin supplementation was employed [10]. On this view, a differentiation stimulus to the target cells would be an unlikely event to happen. In conclusion, there may be a possible role for the remaining plasma fibronectin, as an extra differentiation inductor, which can also modulate the remodeling of the constructed tissue via MMPs activity, in platelet/plasma-based scaffolds used for stem cell differentiation purposes. Future investigators should be aware of
DOI: 10.3109/09537104.2013.836748
this in their future endeavors with these scaffolds for regenerative medicine research protocols.
Acknowledgements We would like to thank Mr Chris Gieseke at University of Texas at San Antonio, for providing excellent assistance in the English language revision of this article. The authors also greatly acknowledge Conselho Nacional de Desenvolvimento Cientı´fico e Tecnolo´gico – CNPq, for a post-doctoral fellowship granted to A.M. (process n. 151248/ 2013-3).
Declaration of interest The author reports no conflict of interest.
References 1. Soto-Gutierrez A, Wertheim JA, Ott HC, Gilbert TW. Perspectives on whole-organ assembly: Moving toward transplantation on demand. J Clin Invest 2012;122:3817–3823. 2. Moroz A, Bittencourt RAC, Almeida RP, Felisbino SL, Deffune E. Platelet lysate 3D scaffold supports mesenchymal stem cell chondrogenesis: An improved approach in cartilage tissue engineering. Platelets 2013;24:219–225. 3. Xie X, Wang Y, Zhao C, Guo S, Liu S, Jia W, Tuan RS, Zhang C. Comparative evaluation of MSCs from bone marrow and adipose tissue seeded in PRP-derived scaffold for cartilage regeneration. Biomaterials 2012;33:7008–7018.
Bioactive scaffolds from plasma and platelets
557
4. Linsley C, Wu B, Tawil B. The effect of fibrinogen, collagen type I, and fibronectin on mesenchymal stem cell growth and differentiation into osteoblasts. Tissue Eng Part A 2013;19: 1416–1423. 5. Moroz A, Delella FK, Lacorte LM, Deffune E, Felisbino SL. Fibronectin induces MMP2 expression in human prostate cancer cells. Biochem Biophys Res Commun 2013;430:1319–1321. 6. Chen Y, Bai B, Zhang S, Ye J, Zhai H, Chen Y, Zhang L, Zeng Y. Study of a novel three-dimensional scaffold to repair bone defect in rabbit. J Biomed Mater Res A 2013. [Epub ahead of print]. doi: 10.1002/jbm.a.34788.. 7. Anitua E, Prado R, Orive G. Endogenous morphogens and fibrin bioscaffolds for stem cell therapeutics. Trends Biotechnol 2013;31: 364–374. 8. Anitua E, Prado R, Azkargorta M, Rodriguez-Sua´rez E, Iloro I, Casado-Vela J, Elortza F, Orive G. High-throughput proteomic characterization of plasma rich in growth factors (PRGF-Endoret)-derived fibrin clot interactome. J Tissue Eng Regen Med 2013. [Epub ahead of print]. doi: 10.1002/ term.1721. 9. Buchta C, Dettke M, Funovics PT, Ho¨cker P, Kno¨bl P, Macher M, Quehenberger P, Treitl C, Worel N. Fibrin sealant produced by the CryoSeal FS System: Product chemistry, material properties and possible preparation in the autologous preoperative setting. Vox Sang 2004;86:257–262. 10. Feng Q, Chai C, Jiang XS, Leong KW, Mao HQ. Expansion of engrafting human hematopoietic stem/progenitor cells in threedimensional scaffolds with surface-immobilized fibronectin. J Biomed Mater Res A 2006;78:781–791.
Copyright of Platelets is the property of Taylor & Francis Ltd and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use.
LETTER
Platelet and plasma bioactive scaffolds for stem cell differentiation: What are we missing? Andrei Moroz1,2, Se´rgio L. Felisbino2, & Elenice Deffune1 1
Cell Engineering Lab, Blood Transfusion Center, Botucatu Medical School, Univ Estadual Paulista, UNESP, Botucatu, SP, Brazil and 2Extracellular Matrix Lab, Department of Morphology, Botucatu Biosciences Institute, Univ Estadual Paulista, UNESP, Botucatu, SP, Brazil
To the editor, Tissue engineering has evolved rapidly over the past 10 years and hold great promise for the treatment of several health injuries, especially due to the accumulated knowledge in stem cell biology, cultivation techniques and differentiation capabilities [1]. Many tissues to be generated in vitro require three dimensional (3D) scaffolds for the successful differentiation of stem cells into the target/desired cell phenotype, such as cartilage tissue and others [1, 2]. Due to this escalating demand, scaffolding technology for cell culture/therapy now allows for more efficient approaches, notably the employment of bioactive materials as their major constituents [2]. One recently published report at this journal demonstrated the feasibility of using platelets as the main component of a bioactive 3D constructed scaffold for mesenchymal stem cell (MSC) cultivation [2]. This scaffold has been found to be suitable for the differentiation of the referred cells in chondrocytes, as demonstrated using extracellular matrix characterization [2] and molecular biology techniques [3]. Moreover, its main component is easy to access, especially in blood transfusion centers. Although this scaffold has been systematically considered to be bioactive largely due to the growth factors, such as PDGF, VEGF, FGF, TGF-b, and others, present at platelets’ granules [3], to date, no one has ever considered a role for plasma fibronectin, as a bioactive agent, which could deliver extra differentiation cues for stem cells cultivated inside these scaffolds. Interestingly, accumulated evidence shows that fibronectin, an abundant protein present at blood plasma, bounded to platelets, and extracellular matrix (ECM), is able to actively modify the biology behavior of both stem [4] and tumoral cells [5]. In this sense, it has been reported that traits such as the activity of matrix metalloproteinases (MMPs), important endopeptidases related to ECM remodeling, which could not be detected at the conditioned medium of prostate normal and tumoral human cells, rapidly increases after short time exposure to fibronectin [5]. Given that the 3D scaffold structure needs to be reabsorbed, as the EMC proteins are produced by the seeded cells, this stimulus on MMP activity could be beneficial during the remodeling of the 3D structure. Moreover, stem cells cultivated at bone differentiation Correspondence: A. Moroz, Univ Estadual Paulista, UNESP, Botucatu Medical School, Blood Transfusion Center, Cell Engineering Lab, District of Rubia˜o Ju´nior S/N, 18618-970, Botucatu, SP, Brazil. Tel: +551438116041 (r 235). E-mail: [email protected]
Keywords Bioactive scaffolds, extracellular matrix, fibronectin, platelet based scaffolds, platelet-rich plasma, stem cells History Received 22 July 2013 Revised 15 August 2013 Accepted 18 August 2013 Published online 31 October 2013
medium exhibited enhanced alkaline phosphatase activity and calcium deposition after exposure to fibronectin coated to culture plates [4], which backs up a possible differentiation role of fibronectin in platelet-derived scaffolds, as a synergistic inductor effect to the stimulus provided from the aforementioned growth factors. Also very relevant, other bioactive scaffolds such as those derived from fibrin sealants are commonly used for stem cell differentiation protocols [6]. For a detailed review on these scaffolds and their applications for regenerative medicine, readers are advised to seek a recently published report [7]. These products, which are commercially available [8, 9] or obtained with an automated device for the production of fibrin sealant [9], have been characterized (nevertheless of the method of obtention) as being enriched in fibronectin content, as observed through the use of specific antibodies and sodium dodecyl sulphate– polyacrylamide gel electrophoresis [9]. Therefore, protocols that employ these scaffolds should also take into consideration the possible influence of fibronectin, on the stimulus provided to the differentiation of the seeded stem cells and their MMPs expression/activity. However, this protein could also exert a proliferative induction of stem cells cultivated in platelet constructed scaffolds, as there are reports of higher proliferation of CD34þ cells when fibronectin supplementation was employed [10]. On this view, a differentiation stimulus to the target cells would be an unlikely event to happen. In conclusion, there may be a possible role for the remaining plasma fibronectin, as an extra differentiation inductor, which can also modulate the remodeling of the constructed tissue via MMPs activity, in platelet/plasma-based scaffolds used for stem cell differentiation purposes. Future investigators should be aware of
DOI: 10.3109/09537104.2013.836748
this in their future endeavors with these scaffolds for regenerative medicine research protocols.
Acknowledgements We would like to thank Mr Chris Gieseke at University of Texas at San Antonio, for providing excellent assistance in the English language revision of this article. The authors also greatly acknowledge Conselho Nacional de Desenvolvimento Cientı´fico e Tecnolo´gico – CNPq, for a post-doctoral fellowship granted to A.M. (process n. 151248/ 2013-3).
Declaration of interest The author reports no conflict of interest.
References 1. Soto-Gutierrez A, Wertheim JA, Ott HC, Gilbert TW. Perspectives on whole-organ assembly: Moving toward transplantation on demand. J Clin Invest 2012;122:3817–3823. 2. Moroz A, Bittencourt RAC, Almeida RP, Felisbino SL, Deffune E. Platelet lysate 3D scaffold supports mesenchymal stem cell chondrogenesis: An improved approach in cartilage tissue engineering. Platelets 2013;24:219–225. 3. Xie X, Wang Y, Zhao C, Guo S, Liu S, Jia W, Tuan RS, Zhang C. Comparative evaluation of MSCs from bone marrow and adipose tissue seeded in PRP-derived scaffold for cartilage regeneration. Biomaterials 2012;33:7008–7018.
Bioactive scaffolds from plasma and platelets
557
4. Linsley C, Wu B, Tawil B. The effect of fibrinogen, collagen type I, and fibronectin on mesenchymal stem cell growth and differentiation into osteoblasts. Tissue Eng Part A 2013;19: 1416–1423. 5. Moroz A, Delella FK, Lacorte LM, Deffune E, Felisbino SL. Fibronectin induces MMP2 expression in human prostate cancer cells. Biochem Biophys Res Commun 2013;430:1319–1321. 6. Chen Y, Bai B, Zhang S, Ye J, Zhai H, Chen Y, Zhang L, Zeng Y. Study of a novel three-dimensional scaffold to repair bone defect in rabbit. J Biomed Mater Res A 2013. [Epub ahead of print]. doi: 10.1002/jbm.a.34788.. 7. Anitua E, Prado R, Orive G. Endogenous morphogens and fibrin bioscaffolds for stem cell therapeutics. Trends Biotechnol 2013;31: 364–374. 8. Anitua E, Prado R, Azkargorta M, Rodriguez-Sua´rez E, Iloro I, Casado-Vela J, Elortza F, Orive G. High-throughput proteomic characterization of plasma rich in growth factors (PRGF-Endoret)-derived fibrin clot interactome. J Tissue Eng Regen Med 2013. [Epub ahead of print]. doi: 10.1002/ term.1721. 9. Buchta C, Dettke M, Funovics PT, Ho¨cker P, Kno¨bl P, Macher M, Quehenberger P, Treitl C, Worel N. Fibrin sealant produced by the CryoSeal FS System: Product chemistry, material properties and possible preparation in the autologous preoperative setting. Vox Sang 2004;86:257–262. 10. Feng Q, Chai C, Jiang XS, Leong KW, Mao HQ. Expansion of engrafting human hematopoietic stem/progenitor cells in threedimensional scaffolds with surface-immobilized fibronectin. J Biomed Mater Res A 2006;78:781–791.
Copyright of Platelets is the property of Taylor & Francis Ltd and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use.
