Biological activity of laminin peptide-conjugated alginate and chitosan matrices.

Laminin active peptide-conjugated chitosan mambranes have been previously demonstrated as a useful biomaterial for tissue engineering. Here, three laminin active peptides, A99 (AGTFALRGDNPQG), AG73 (RKRLQVQLSIRT), and EF1zz (ATLQLQEGRLHFXFDLGKGR, X: Nle), which interact with integrin αvβ3, syndecans, and integrin α2β1, respectively, were conjugated to alginate and evaluated the biological activities. A99-alginate (3-3000 ng/mm(2)) promoted cell attachment depending on the amount of alginate. More than 300 ng/mm(2) of the A99-alginate matrices effectively promoted cell attachment, cell spreading with well-organized actin stress fibers, and neurite outgrowth. AG73- and EF1zz-alginates promoted strong cell attachment at the all amounts (3-3000 ng/mm(2)). A99-alginate (30-3000 ng/mm(2)) promoted strong neurite outgrowth but lower amounts of A99-alginate (3 ng/mm(2)) showed weak activity. In contrast, AG73-alginates (3-30 ng/mm(2)) showed strong neurite outgrowth activity but higher amounts of AG73-alginate (300-3000 ng/mm(2)) decreased the activity. These data indicate that neurite outgrowth activity of peptide-alginate matrices is peptide specific and the activity is dependent on the amount of alginate. Further, biological activities of the peptides on alginate and chitosan matrices were different, suggesting that the integrin- and syndecan-mediated cellular functions on the peptide-matrices are highly influenced by the scaffold structure including polysaccharide types and amounts. The laminin active peptide-conjugated alginate and chitosan matrices can control receptor type specific functions and are useful for tissue engineering.

[1]  M. Nomizu,et al.  Functional Sites in the Laminin Alpha Chains , 2005, Connective tissue research.

[2]  Horst Kessler,et al.  RGD modified polymers: biomaterials for stimulated cell adhesion and beyond. , 2003, Biomaterials.

[3]  R. Timpl Macromolecular organization of basement membranes. , 1996, Current opinion in cell biology.

[4]  M. Jasionowski,et al.  Injectable gels for tissue engineering , 2001, The Anatomical record.

[5]  J. Miner,et al.  Laminin functions in tissue morphogenesis. , 2004, Annual review of cell and developmental biology.

[6]  Richard O Hynes,et al.  Integrins Bidirectional, Allosteric Signaling Machines , 2002, Cell.

[7]  S. Yamashina,et al.  A novel cell‐adhesive scaffold material for delivering keratinocytes reduces granulation tissue in dermal wounds , 2009, Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society.

[8]  Y. Suzuki,et al.  In vivo evaluation of a novel alginate dressing. , 1999, Journal of biomedical materials research.

[9]  M. Nomizu,et al.  Mixed peptide-chitosan membranes to mimic the biological activities of a multifunctional laminin alpha1 chain LG4 module. , 2009, Biomaterials.

[10]  C. Laurencin,et al.  Biologically active chitosan systems for tissue engineering and regenerative medicine. , 2008, Current topics in medicinal chemistry.

[11]  H. Kleinman,et al.  Cell Type-specific Differences in Glycosaminoglycans Modulate the Biological Activity of a Heparin-binding Peptide (RKRLQVQLSIRT) from the G Domain of the Laminin α1 Chain* , 2001, The Journal of Biological Chemistry.

[12]  D J Mooney,et al.  Alginate hydrogels as synthetic extracellular matrix materials. , 1999, Biomaterials.

[13]  S. Ogata,et al.  A novel covalently crosslinked gel of alginate and silane with the ability to form bone-like apatite. , 2004, Journal of biomedical materials research. Part A.

[14]  Smadar Cohen,et al.  Optimization of cardiac cell seeding and distribution in 3D porous alginate scaffolds. , 2002, Biotechnology and bioengineering.

[15]  Mehrdad Hamidi,et al.  Hydrogel nanoparticles in drug delivery. , 2008, Advanced drug delivery reviews.

[16]  A. Utani,et al.  Biological Activities of Homologous Loop Regions in the Laminin α Chain G Domains* , 2003, Journal of Biological Chemistry.

[17]  H. Kleinman,et al.  Matrigel: basement membrane matrix with biological activity. , 2005, Seminars in cancer biology.

[18]  P. Ma,et al.  Ionically crosslinked alginate hydrogels as scaffolds for tissue engineering: part 1. Structure, gelation rate and mechanical properties. , 2001, Biomaterials.

[19]  Stephen F Badylak,et al.  The extracellular matrix as a biologic scaffold material. , 2007, Biomaterials.

[20]  C. Renner,et al.  Synthetic heterotrimeric collagen peptides as mimics of cell adhesion sites of the basement membrane. , 2004, Biopolymers.

[21]  J. Hubbell,et al.  Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering , 2005, Nature Biotechnology.

[22]  A. Utani,et al.  Identification of Cell Binding Sites in the Laminin α1 Chain Carboxyl-terminal Globular Domain by Systematic Screening of Synthetic Peptides (*) , 1995, The Journal of Biological Chemistry.

[23]  H. Kleinman,et al.  Angiogenic activity of syndecan-binding laminin peptide AG73 (RKRLQVQLSIRT). , 2007, Archives of biochemistry and biophysics.

[24]  Taku Sato,et al.  Laminin‐1 peptide‐conjugated chitosan membranes as a novel approach for cell engineering , 2003, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[25]  M. Wheatley,et al.  Microencapsulated liposomes in controlled drug delivery: strategies to modulate drug release and eliminate the burst effect. , 2003, Journal of pharmaceutical sciences.

[26]  G. Cardenas,et al.  Chitosan composite films. Biomedical applications , 2008, Journal of materials science. Materials in medicine.

[27]  H. Kleinman,et al.  Cell Adhesive Sequences in Mouse Laminin β1 Chain , 2000 .

[28]  C. Chiu,et al.  Development of two alginate-based wound dressings , 2008, Journal of materials science. Materials in medicine.

[29]  H. Kleinman,et al.  Integrin-dependent cell behavior on ECM peptide-conjugated chitosan membranes. , 2007, Biopolymers.

[30]  S. Yamashina,et al.  Laminin peptide-conjugated chitosan membrane: Application for keratinocyte delivery in wounded skin. , 2006, Journal of biomedical materials research. Part A.

[31]  R V Bellamkonda,et al.  The influence of physical structure and charge on neurite extension in a 3D hydrogel scaffold. , 1998, Journal of biomaterials science. Polymer edition.

[32]  Hiroshi Morioka,et al.  Syndecan Binding Sites in the Laminin α1 Chain G Domain , 2003 .

[33]  H. Kleinman,et al.  Cell Binding Sequences in Mouse Laminin α1 Chain* , 1998, The Journal of Biological Chemistry.

[34]  Mina J Bissell,et al.  Modeling tissue-specific signaling and organ function in three dimensions , 2003, Journal of Cell Science.

[35]  H. Kleinman,et al.  Laminin-1 and Laminin-2 G-domain Synthetic Peptides Bind Syndecan-1 and Are Involved in Acinar Formation of a Human Submandibular Gland Cell Line* , 1998, The Journal of Biological Chemistry.

[36]  J. Boateng,et al.  Wound healing dressings and drug delivery systems: a review. , 2008, Journal of pharmaceutical sciences.

[37]  J. Suh,et al.  Application of chitosan-based polysaccharide biomaterials in cartilage tissue engineering: a review. , 2000, Biomaterials.

[38]  B. Nies,et al.  Surface Coating with Cyclic RGD Peptides Stimulates Osteoblast Adhesion and Proliferation as well as Bone Formation , 2000, Chembiochem : a European journal of chemical biology.

[39]  H. Kleinman,et al.  Identification of Cell Binding Sequences in Mouse Laminin γ1 Chain by Systematic Peptide Screening* , 1997, The Journal of Biological Chemistry.