Corneal Stromal Cell Growth on Gelatin/Chondroitin Sulfate Scaffolds Modified at Different NHS/EDC Molar Ratios
Tóm tắt
A nanoscale modification strategy that can incorporate chondroitin sulfate (CS) into the cross-linked porous gelatin materials has previously been proposed to give superior performance for designed corneal keratocyte scaffolds. The purpose of this work was to further investigate the influence of carbodiimide chemistry on the characteristics and biofunctionalities of gelatin/CS scaffolds treated with varying N-hydroxysuccinimide (NHS)/1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide hydrochloride (EDC) molar ratios (0-1) at a constant EDC concentration of 10 mM. Results of Fourier transform infrared spectroscopy and dimethylmethylene blue assays consistently indicated that when the NHS to EDC molar ratio exceeds a critical level (i.e., 0.5), the efficiency of carbodiimide-mediated biomaterial modification is significantly reduced. With the optimum NHS/EDC molar ratio of 0.5, chemical treatment could achieve relatively high CS content in the gelatin scaffolds, thereby enhancing the water content, glucose permeation, and fibronectin adsorption. Live/Dead assays and interleukin-6 mRNA expression analyses demonstrated that all the test samples have good cytocompatibility without causing toxicity and inflammation. In the molar ratio range of NHS to EDC from 0 to 0.5, the cell adhesion ratio and proliferation activity on the chemically modified samples significantly increased, which is attributed to the increasing CS content. Additionally, the materials with highest CS content (0.143 ± 0.007 nmol/10 mg scaffold) showed the greatest stimulatory effect on the biosynthetic activity of cultivated keratocytes. These findings suggest that a positive correlation is noticed between the NHS to EDC molar ratio and the CS content in the biopolymer matrices, thereby greatly affecting the corneal stromal cell growth.
Từ khóa
Tài liệu tham khảo
Lai, 2007, Functional biomedical polymers for corneal regenerative medicine, React. Funct. Polym, 67, 1284, 10.1016/j.reactfunctpolym.2007.07.060
Li, 2003, Cellular and nerve regeneration within a biosynthetic extracellular matrix for corneal transplantation, Proc. Natl. Acad. Sci. USA, 100, 15346, 10.1073/pnas.2536767100
Liu, 2006, A simple, cross-linked collagen tissue substitute for corneal implantation, Invest. Ophthalmol. Vis. Sci, 47, 1869, 10.1167/iovs.05-1339
Vrana, 2008, Development of a reconstructed cornea from collagen-chondroitin sulfate foams and human cell cultures, Invest. Ophthalmol. Vis. Sci, 49, 5325, 10.1167/iovs.07-1599
Lai, 2010, Ocular biocompatibility of carbodiimide cross-linked hyaluronic acid hydrogels for cell sheet delivery carriers, J. Biomater. Sci. Polym. Ed, 21, 359, 10.1163/156856209X416980
Lai, 2010, Functional assessment of cross-linked porous gelatin hydrogels for bioengineered cell sheet carriers, Biomacromolecules, 11, 1387, 10.1021/bm100213f
Ma, 2010, Carbodiimide cross-linked amniotic membranes for cultivation of limbal epithelial cells, Biomaterials, 31, 6647, 10.1016/j.biomaterials.2010.05.034
Nakajima, 1995, Mechanism of amide formation by carbodiimide for bioconjugation in aqueous media, Bioconjug. Chem, 6, 123, 10.1021/bc00031a015
Lai, 2012, A gelatin-g-poly(N-isopropylacrylamide) biodegradable in situ gelling delivery system for the intracameral administration of pilocarpine, Biomaterials, 33, 2372, 10.1016/j.biomaterials.2011.11.085
Lai, 2012, Nanoscale modification of porous gelatin scaffolds with chondroitin sulfate for corneal stromal tissue engineering, Int. J. Nanomed, 7, 1101, 10.2147/IJN.S28753
Wang, 2009, Applications and degradation of proteins used as tissue engineering materials, Materials, 2, 613, 10.3390/ma2020613
Kuijpers, 2000, Cross-linking and characterisation of gelatin matrices for biomedical applications, J. Biomater. Sci. Polym. Ed, 11, 225, 10.1163/156856200743670
Dijkstra, 1996, Cross-linking of dermal sheep collagen using a water-soluble carbodiimide, Biomaterials, 17, 765, 10.1016/0142-9612(96)81413-X
Lu, 2008, A methodology based on the “anterior chamber of rabbit eyes” model for noninvasively determining the biocompatibility of biomaterials in an immune privileged site, J. Biomed. Mater. Res. A, 86, 108, 10.1002/jbm.a.31619
Hsiue, 2006, A novel strategy for corneal endothelial reconstruction with a bioengineered cell sheet, Transplantation, 81, 473, 10.1097/01.tp.0000194864.13539.2c
Lai, 2007, Tissue-engineered human corneal endothelial cell sheet transplantation in a rabbit model using functional biomaterials, Transplantation, 84, 1222, 10.1097/01.tp.0000287336.09848.39
Malafaya, 2007, Natural-origin polymers as carriers and scaffolds for biomolecules and cell delivery in tissue engineering applications, Adv. Drug Deliv. Rev, 59, 207, 10.1016/j.addr.2007.03.012
Leonard, 1997, Refractive indices of the collagen fibrils and extrafibrillar material of the corneal stroma, Biophys. J, 72, 1382, 10.1016/S0006-3495(97)78784-8
Monti, 2002, Increased corneal hydration induced by potential ocular penetration enhancers: assessment by differential scanning calorimetry (DSC) and by desiccation, Int. J. Pharm, 232, 139, 10.1016/S0378-5173(01)00907-3
Hsiue, 2002, Absorbable sandwich-like membrane for retinal-sheet transplantation, J. Biomed. Mater. Res, 61, 19, 10.1002/jbm.2000
Lai, 2009, Low Bloom strength gelatin as a carrier for potential use in retinal sheet encapsulation and transplantation, Biomacromolecules, 10, 310, 10.1021/bm801039n
Lu, 2008, Carbodiimide cross-linked hyaluronic acid hydrogels as cell sheet delivery vehicles: characterization and interaction with corneal endothelial cells, J. Biomater. Sci.-Polym. Ed, 19, 1, 10.1163/156856208783227695
Lai, 2012, Solvent composition is critical for carbodiimide cross-linking of hyaluronic acid as an ophthalmic biomaterial, Materials, 5, 1986, 10.3390/ma5101986
Zeugolis, 2009, Cross-linking of extruded collagen fibers—a biomimetic three-dimensional scaffold for tissue engineering applications, J. Biomed. Mater. Res. A, 89, 895, 10.1002/jbm.a.32031
Jayasuriya, 2003, Piezoelectric and mechanical properties in bovine cornea, J. Biomed. Mater. Res. A, 66, 260, 10.1002/jbm.a.10536
Chayakul, 1982, The enzymatic activities in the alkali-burnt rabbit cornea, Graefes Arch. Clin. Exp. Ophthalmol, 218, 145, 10.1007/BF02215652
Lai, 2010, Biocompatibility of chemically cross-linked gelatin hydrogels for ophthalmic use, J. Mater. Sci.-Mater. Med, 21, 1899, 10.1007/s10856-010-4035-3
Lai, 2010, In vitro response of retinal pigment epithelial cells exposed to chitosan materials prepared with different cross-linkers, Int. J. Mol. Sci, 11, 5256, 10.3390/ijms11125256
Lai, 2012, Synthesis, characterization and ocular biocompatibility of potential keratoprosthetic hydrogels based on photopolymerized poly(2-hydroxyethyl methacrylate)-co-poly(acrylic acid), J. Mater. Chem, 22, 1812, 10.1039/C1JM14211A
Lai, 2012, Biocompatibility of genipin and glutaraldehyde cross-linked chitosan materials in the anterior chamber of the eye, Int. J. Mol. Sci, 13, 10970, 10.3390/ijms130910970
Strehin, 2010, A versatile pH sensitive chondroitin sulfate-PEG tissue adhesive and hydrogel, Biomaterials, 31, 2788, 10.1016/j.biomaterials.2009.12.033
Yamada, 1978, Fibronectins-adhesive glycoproteins of cell surface and blood, Nature, 275, 179, 10.1038/275179a0
Wasylnka, 2000, Adhesion of Aspergillus species to extracellular matrix proteins: evidence for involvement of negatively charged carbohydrates on the conidial surface, Infect. Immun, 68, 3377, 10.1128/IAI.68.6.3377-3384.2000
Lai, 2012, Adhesion, phenotypic expression, and biosynthetic capacity of corneal keratocytes on surfaces coated with hyaluronic acid of different molecular weights, Acta Biomater, 8, 1068, 10.1016/j.actbio.2011.11.012
Rammelt, 2006, Coating of titanium implants with collagen, RGD peptide and chondroitin sulfate, Biomaterials, 27, 5561, 10.1016/j.biomaterials.2006.06.034
Zou, 2009, Specific interactions between human fibroblasts and particular chondroitin sulfate molecules for wound healing, Acta Biomater, 5, 1588, 10.1016/j.actbio.2008.12.001
Milev, 1998, The core protein of the chondroitin sulfate proteoglycan phosphacan is a high-affinity ligand of fibroblast growth factor-2 and potentiates its mitogenic activity, J. Biol. Chem, 273, 21439, 10.1074/jbc.273.34.21439
Smith, 2007, Heparan and chondroitin sulfate on growth plate perlecan mediate binding and delivery of FGF-2 to FGF receptors, Matrix Biol, 26, 175, 10.1016/j.matbio.2006.10.012
Pieper, 2001, Linkage of chondroitin-sulfate to type I collagen scaffolds stimulates the bioactivity of seeded chondrocytes in vitro, Biomaterials, 22, 2359, 10.1016/S0142-9612(00)00423-3
Bassleer, 1998, Effects of chondroitin sulfate and interleukin-1β on human articular chondrocytes cultivated in clusters, Osteoarthritis Cartilage, 6, 196, 10.1053/joca.1998.0112
Cao, 2008, EDC/NHS-crosslinked type II collagen-chondroitin sulfate scaffold: characterization and in vitro evaluation, J. Mater. Sci.-Mater. Med, 19, 567, 10.1007/s10856-007-3281-5
Lai, 2011, Influence of cross-linker concentration on the functionality of carbodiimide cross-linked gelatin membranes for retinal sheet carriers, J. Biomater. Sci.-Polym. Ed, 22, 277, 10.1163/092050609X12603600753204
Lai, 2006, Effect of charge and molecular weight on the functionality of gelatin carriers for corneal endothelial cell therapy, Biomacromolecules, 7, 1836, 10.1021/bm0601575
Lai, 2010, Evaluation of cross-linked gelatin membranes as delivery carriers for retinal sheets, Mater. Sci. Eng. C, 30, 677, 10.1016/j.msec.2010.02.024
Lai, 2011, Evaluation of cross-linking time for porous gelatin hydrogels on cell sheet delivery performance, J. Mech. Med. Biol, 11, 967, 10.1142/S0219519411004873
Lai, 2006, Bioengineered human corneal endothelium for transplantation, Arch. Ophthalmol, 124, 1441, 10.1001/archopht.124.10.1441
Lai, J.Y. The role of bloom index of gelatin on the interaction with retinal pigment epithelial cells.