Restricted accessResearch articleFirst published online 2015-1
In vitro and in vivo evaluation of a novel collagen/cellulose nanocrystals scaffold for achieving the sustained release of basic fibroblast growth factor
Tissue-engineered dermis is thought to be the best treatment for skin defects; however, slow vascularization of these biomaterial scaffolds limits their clinical application. Exogenous administration of angiogenic growth factors is highly desirable for tissue regeneration. In this study, biodegradable gelatin microspheres (GMs) containing basic fibroblast growth factor (bFGF) were fabricated and incorporated into a porous collagen/cellulose nanocrystals (CNCs) scaffold, as a platform for long-term release and consequent angiogenic boosting. The physicochemical properties of these scaffolds were examined and the in vitro release pattern of bFGF from scaffolds was measured by ELISA. Collagen/CNCs scaffolds with and without bFGF-GMs were incubated with human umbilical vein endothelial cells for 1 week, results showed that the scaffolds with bFGF-GMs significantly augmented cell proliferation. Then, four different groups of scaffolds were implanted subcutaneously into Sprague–Dawley rats to study angiogenesis in vivo via macroscopic observation, and hematoxylin and eosin and immunohistochemical staining. The results suggested that the collagen/CNCs/bFGF-GMs scaffolds had a significantly higher number of newly formed and mature blood vessels, and the fastest degradation rate. This study demonstrated that collagen/CNCs/bFGF-GMs scaffolds have great potential in skin tissue engineering.
MooneyDJMikosAG. Growing new organs. Scient Am1999; 280: 60–67.
2.
PieperJHafmansTVan WachemP. Loading of collagen–heparan sulfate matrices with bFGF promotes angiogenesis and tissue generation in rats. J Biomed Mater Res2002; 62: 185–194.
3.
BaigueraSRibattiD. Endothelialization approaches for viable engineered tissues. Angiogenesis2013; 16: 1–14.
4.
O’BrienFJHarleyBYannasIV. The effect of pore size on cell adhesion in collagen-GAG scaffolds. Biomaterials2005; 26 : 433–441.
5.
RickertDMosesMLendleinA. The importance of angiogenesis in the interaction between polymeric biomaterials and surrounding tissue. Clin Hemorheol Micro2003; 28: 175–181.
JungebluthPBaderABaigueraS. The concept of in vivo airway tissue engineering. Biomaterials2012; 33: 4319–4326.
8.
CaoRBråkenhielmEPawliukR. Angiogenic synergism, vascular stability and improvement of hind-limb ischemia by a combination of PDGF-BB and FGF-2. Nat Med2003; 9: 604–613.
9.
EhrbarMZeisbergerSMRaeberGP. The role of actively released fibrin-conjugated VEGF for VEGF receptor 2 gene activation and the enhancement of angiogenesis. Biomaterials2008; 29: 1720–1729.
10.
NillesenSGeutjesPJWismansR. Increased angiogenesis and blood vessel maturation in acellular collagen-heparin scaffolds containing both FGF2 and VEGF. Biomaterials2007; 28: 1123–1131.
11.
RichardsonTPPetersMCEnnettAB. Polymeric system for dual growth factor delivery. Nat Biotechnol2001; 19: 1029–1034.
12.
UchiHIgarashiAUrabeK. Clinical efficacy of basic fibroblast growth factor (bFGF) for diabetic ulcer. Eur J Dermatol2009; 19: 461–468.
13.
KawaiKSuzukiSTabataY. Accelerated tissue regeneration through incorporation of basic fibroblast growth factor-impregnated gelatin microspheres into artificial dermis. Biomaterials2000; 21: 489–499.
14.
MuneuchiGSuzukiSMoriueT. Combined treatment using artificial dermis and basic fibroblast growth factor (bFGF) for intractable fingertip ulcers caused by atypical burn injuries. Burns2005; 31: 514–517.
15.
ItoKItoSSekineM. Reconstruction of the soft tissue of a deep diabetic foot wound with artificial dermis and recombinant basic fibroblast growth factor. Plast Reconstr Surg2005; 115: 567–572.
16.
AkitaSAkinoKTanakaK. A basic fibroblast growth factor improves lower extremity wound healing with a porcine-derived skin substitute. J Trauma2008; 64: 809–815.
17.
KandaNMorimotoNAyvazyanAA. Evaluation of a novel collagen-gelatin scaffold for achieving the sustained release of basic fibroblast growth factor in a diabetic mouse model. J Tissue Eng Regen Med2014; 8: 29–40.
18.
ZhuXHTabataYWangCH. Delivery of basic fibroblast growth factor from gelatin microsphere scaffold for the growth of human umbilical vein endothelial cells. Tissue Eng A2008; 14: 1939–1947.
19.
Del GaudioCBaigueraSBoieriM. Induction of angiogenesis using VEGF releasing genipin-crosslinked electrospun gelatin mats. Biomaterials2013; 34: 7754–7765.
20.
LiWGuoRLanY. Preparation and properties of cellulose nanocrystals reinforced collagen composite films. J Biomed Mater Res A2014; 102A: 1131–1139.
21.
TakahashiYYamamotoMTabataY. Osteogenic differentiation of mesenchymal stem cells in biodegradable sponges composed of gelatin and beta-tricalcium phosphate. Biomaterials2005; 26: 3587–3596.
22.
SpizzirriUIemmaFAltimariI. Grafted gelatin microspheres as potential pH-responsive devices. J Mater Sci2012; 47: 3648–3657.
23.
d’AngeloIOlivieroOUngaroF. Engineering strategies to control vascular endothelial growth factor stability and levels in a collagen matrix for angiogenesis: The role of heparin sodium salt and the PLGA-based microsphere approach. Acta Biomater2013; 9: 7389–7398.
24.
BorselliCUngaroFOlivieroO. Bioactivation of collagen matrices through sustained VEGF release from PLGA microspheres. J Biomed Mater Res A2010; 92: 94–102.
25.
MaLGaoCMaoZ. Collagen/chitosan porous scaffolds with improved biostability for skin tissue engineering. Biomaterials2003; 24: 4833–4841.
26.
DaiN-TWilliamsonMKhammoN. Composite cell support membranes based on collagen and polycaprolactone for tissue engineering of skin. Biomaterials2004; 25: 4263–4271.
27.
HiraokaYKimuraYUedaH. Fabrication and biocompatibility of collagen sponge reinforced with poly (glycolic acid) fiber. Tissue Eng2003; 9: 1101–1112.
28.
PooyanPTannenbaumRGarmestaniH. Mechanical behavior of a cellulose-reinforced scaffold in vascular tissue engineering. J Mech Behav Biomed2012; 7 : 50–59.
29.
Matos RuizMCavailleJDufresneA. Processing and characterization of new thermoset nanocomposites based on cellulose whiskers. Compos Interf2000; 7: 117–131.
30.
TabataYIkadaY. Protein release from gelatin matrices. Adv Drug Deliv Rev1998; 31: 287–301.
31.
LiuHFanHCuiY. Effects of the controlled-released basic fibroblast growth factor from chitosan-gelatin microspheres on human fibroblasts cultured on a chitosan-gelatin scaffold. Biomacromolecules2007; 8: 1446–1455.
32.
KawaiKSuzukiSTabataY. Accelerated tissue regeneration through incorporation of basic fibroblast growth factor-impregnated gelatin microspheres into artificial dermis. Biomaterials2000; 21: 489–499.
33.
DongXMRevolJ-fGrayDG. Effect of microcrystallite preparation conditions on the formation of colloid crystals of cellulose. Cellulose1998; 5: 19–32.
34.
LiuYMaLGaoC. Facile fabrication of the glutaraldehyde cross-linked collagen/chitosan porous scaffold for skin tissue engineering. Mater Sci Eng C2012; 32: 2361–2366.
35.
LeeJEKimKEKwonIC. Effects of the controlled-released TGF-beta 1 from chitosan microspheres on chondrocytes cultured in a collagen/chitosan/glycosaminoglycan scaffold. Biomaterials2004; 25: 4163–4173.
36.
ZhouCWuQYueY. Application of rod-shaped cellulose nanocrystals in polyacrylamide hydrogels. J Colloid Interf Sci2011; 353: 116–123.
37.
KimSKangYKruegerCA. Sequential delivery of BMP-2 and IGF-1 using a chitosan gel with gelatin microspheres enhances early osteoblastic differentiation. Acta Biomater2012; 8: 1768–1777.
38.
AbitbolTJohnstoneTQuinnTM. Reinforcement with cellulose nanocrystals of poly (vinyl alcohol) hydrogels prepared by cyclic freezing and thawing. Soft Matter2011; 7: 2373–2379.
39.
BalakrishnanBMohantyMUmashankarP. Evaluation of an in situ forming hydrogel wound dressing based on oxidized alginate and gelatin. Biomaterials2005; 26: 6335–6342.
40.
ChoiYSHongSRLeeYM. Study on gelatin-containing artificial skin: I. Preparation and characteristics of novel gelatin-alginate sponge. Biomaterials1999; 20: 409–417.
