Polymeric matrices mimicking multiple functions of the ECM are expected to enable a material induced regeneration of tissues. Here, we investigated the adipogenic differentiation of human adipose derived mesenchymal stem cells (hADSCs) in a 3D architectured gelatin based hydrogel (ArcGel) prepared from gelatin and L-lysine diisocyanate ethyl ester (LDI) in an one-step process, in which the formation of an open porous morphology and the chemical network formation were integrated. The ArcGel was designed to support adipose tissue regeneration with its 3D porous structure, high cell biocompatibility, and mechanical properties compatible with human subcutaneous adipose tissue. The ArcGel could support initial cell adhesion and survival of hADSCs. Under static culture condition, the cells could migrate into the inner part of the scaffold with a depth of 840±120 μm after 4 days, and distributed in the whole scaffold (2 mm in thickness) within 14 days. The cells proliferated in the scaffold and the fold increase of cell number after 7 days of culture was 2.55±0.08. The apoptotic rate of hADSCs in the scaffold was similar to that of cells maintained on tissue culture plates. When cultured in adipogenic induction medium, the hADSCs in the scaffold differentiated into adipocytes with a high efficiency (93±1%). Conclusively, this gelatin based 3D scaffold presented high cell compatibility for hADSC cultivation and differentiation, which could serve as a potential implant material in clinical applications for adipose tissue reparation and regeneration.
BuckyL.P. and PercecI., The science of autologous fat grafting: Views on current and future approaches to neoadipogenesis, Aesthet Surg J28 (2008), 313–321; quiz 322-324.
TabitC.J., SlackG.C., FanK., WanD.C. and BradleyJ.P., Fat grafting versus adipose-derived stem cell therapy: Distinguishing indications, techniques, and outcomes, Aesthetic Plast Surg36 (2012), 704–713.
4.
TremoladaC., PalmieriG. and RicordiC., Adipocyte transplantation and stem cells: Plastic surgery meets regenerative medicine, Cell Transplant19 (2010), 1217–1223.
5.
AndersonJ.M., RodriguezA. and ChangD.T., Foreign body reaction to biomaterials, Semin Immunol20 (2008), 86–100.
6.
YoungD.A. and ChristmanK.L., Injectable biomaterials for adipose tissue engineering, Biomed Mater7 (2012), 024104.
7.
FranzS., RammeltS., ScharnweberD. and SimonJ.C., Immune responses to implants – A review of the implications for the design of immunomodulatory biomaterials, Biomaterials32 (2011), 6692–6709.
8.
LeviB., JamesA.W., GlotzbachJ.P., WanD.C., CommonsG.W. and LongakerM.T., Depot-specific variation in the osteogenic and adipogenic potential of human adipose-derived stromal cells, Plast Reconstr Surg126 (2010), 822–834.
9.
RodriguezA.M., ElabdC., AmriE.Z., AilhaudG. and DaniC., The human adipose tissue is a source of multipotent stem cells, Biochimie87 (2005), 125–128.
10.
HongL., PeptanI.A., ColpanA. and DawJ.L., Adipose tissue engineering by human adipose-derived stromal cells, Cells Tissues Organs183 (2006), 133–140.
11.
KimW.S., ParkB.S. and SungJ.H., Protective role of adipose-derived stem cells and their soluble factors in photoaging, Arch Dermatol Res301 (2009), 329–336.
12.
SalgadoA.J., ReisR.L., SousaN. and GimbleJ.M., Adipose tissue derived stem cells secretome: Soluble factors and their roles in regenerative medicine, Curr Stem Cell Res T5 (2010), 103–110.
13.
KakudoN., ShimotsumaA. and KusumotoK., Fibroblast growth factor-2 stimulates adipogenic differentiation of human adipose-derived stem cells, Biochem Bioph Res Co359 (2007), 239–244.
14.
Cosgriff-HernandezE. and MikosA.G., New biomaterials as scaffolds for tissue engineering, Pharm Res25 (2008), 2345–2347.
15.
RahamanM.N. and MaoJ.J., Stem cell-based composite tissue constructs for regenerative medicine, Biotechnol Bioeng91 (2005), 261–284.
16.
MorganS.M., AinsworthB.J., KanczlerJ.M., BabisterJ.C., ChaudhuriJ.B. and OreffoR.O.C., Formation of a human-derived fat tissue layer in P(DL)LGA hollow fibre scaffolds for adipocyte tissue engineering, Biomaterials30 (2009), 1910–1917.
17.
WangX.H., SuiS.C., YanY.N. and ZhangR.J., Design and Fabrication of PLGA Sandwiched Cell/Fibrin Constructs for Complex Organ Regeneration, J Bioact Compat Polym25 (2010), 229–240.
18.
YeonB., ParkM.H., MoonH.J., KimS.J., CheonY.W. and JeongB., 3D Culture of adipose-tissue-derived stem cells mainly leads to chondrogenesis in poly(ethylene glycol)-poly(L-alanine) diblock copolymer thermogel, Biomacromolecules14 (2013), 3256–3266.
19.
ZambonJ.P., BarrettoL.S.D., NakamuraA.N.S.E., DuailibiS., LeiteK., MagalhaesR.S., OrlandoG., RossC.L., PelosoA. and AlmeidaF.G., Histological changes induced by Polyglycolic-Acid (PGA) scaffolds seeded with autologous adipose or muscle-derived stem cells when implanted on rabbit bladder, Organogenesis10 (2014), 278–288.
20.
CasadeiA., EpisR., FerroniL., ToccoI., GardinC., BressanE., SivolellaS., VindigniV., PintonP., MucciG. and ZavanB., Adipose tissue regeneration: A state of the art, J Biomed Biotechnol (2012), 462543.
21.
ChoiJ.H., GimbleJ.M., LeeK., MarraK.G., RubinJ.P., YooJ.J., Vunjak-NovakovicG. and KaplanD.L., Adipose tissue engineering for soft tissue regeneration, Tiss Eng, Part B16 (2010), 413–426.
22.
RubinJ.P., BennettJ.M., DoctorJ.S., TebbetsB.M. and MarraK.G., Collagenous microbeads as a scaffold for tissue engineering with adipose-derived stem cells, Plast Reconstr Surg120 (2007), 414–424.
23.
MauneyJ.R., NguyenT., GillenK., Kirker-HeadC., GimbleJ.M. and KaplanD.L., Engineering adipose-like tissue in vitro and in vivo utilizing human bone marrow and adipose-derived mesenchymal stem cells with silk fibroin 3D scaffolds, Biomaterials28 (2007), 5280–5290.
24.
ItoiY., TakatoriM., HyakusokuH. and MizunoH., Comparison of readily available scaffolds for adipose tissue engineering using adipose-derived stem cells, J Plast Reconstr Aes63 (2010), 858–864.
25.
RoseJ.B., PacelliS., Haj ElA.J., DuaH.S., HopkinsonA., WhiteL.J. and RoseF.R.A.J., Gelatin-based materials in ocular tissue engineering, Materials7 (2014), 3106–3135.
26.
NeffeA.T., PierceB.F., TronciG., MaN., PittermannE., GebauerT., FrankO., SchossigM., XuX., WillieB.M., FornerM., EllinghausA., LienauJ., DudaG.N. and LendleinA., One step creation of multifunctional 3D architectured hydrogels inducing bone regeneration, Adv Mater27 (2015), 1738–1744.
27.
XuX., WangW.W., KratzK., FangL., LiZ.D., KurtzA., MaN. and LendleinA., Controlling major cellular processes of human mesenchymal stem cells using microwell structures, Adv Healthc Mater3 (2014), 1991–2003.
28.
AldridgeA., KouroupisD., ChurchmanS., EnglishA., InghamE. and JonesE., Assay validation for the assessment of adipogenesis of multipotential stromal cells–a direct comparison of four different methods, Cytotherapy15 (2013), 89–101.
29.
NagahamaH., RaniV.V.D., ShalumonK.T., JayakumarR., NairS.V., KoiwaS., FuruikeT. and TamuraH., Preparation, characterization, bioactive and cell attachment studies of alpha-chitin/gelatin composite membranes, Int J Biol Macromol44 (2009), 333–337.
30.
DavidenkoN., SchusterC.F., BaxD.V., FarndaleR.W., HamaiaS., BestS.M., CameronR.E., Evaluation of cell binding to collagen and gelatin: A study of the effect of 2D and 3D architecture and surface chemistry, J Mater Sci-Mater M27 (2016).
31.
BigiA., CojazziG., PanzavoltaS., RubiniK. and RoveriN., Mechanical and thermal properties of gelatin films at different degrees of glutaraldehyde crosslinking, Biomaterials22 (2001), 763–768.
32.
LienS.M., LiW.T. and HuangT.J., Genipin-crosslinked gelatin scaffolds for articular cartilage tissue engineering with a novel crosslinking method, Mater Sci Eng, C28 (2008), 36–43.
33.
ChangJ.Y., LinJ.H., YaoC.H., ChenJ.H., LaiT.Y. and ChenY.S., In vivo evaluation of a biodegradable EDC/NHS-cross-linked gelatin peripheral nerve guide conduit material, Macromol Biosci7 (2007), 500–507.
34.
TronciG., NeffeA.T., PierceB.F. and LendleinA., An entropy-elastic gelatin-based hydrogel system, J Mater Chem20 (2010), 8875–8884.
35.
MajnoG. and JorisI., Apoptosis, oncosis, and necrosis - an overview of cell-death, Am J Pathol146 (1995), 3–15.
36.
KerrJ.F., WyllieA.H. and CurrieA.R., Apoptosis: A basic biological phenomenon with wide-ranging implications in tissue kinetics, Br J Cancer26 (1972), 239–257.
GobaaS., HoehnelS. and LutolfM.P., Substrate elasticity modulates the responsiveness of mesenchymal stem cells to commitment cues, Integr Biol7 (2015), 1135–1142.
39.
PlaczekM.R., ChungI.M., MacedoH.M., IsmailS., BlancoT.M., LimM., ChaJ.M., FauziI., KangY., YeoD.C.L., MaC.Y.J., PolakJ.M., PanoskaltsisN. and MantalarisA., Stem cell bioprocessing: Fundamentals and principles, J Royal Soc Interf6 (2009), 209–232.
40.
PolakD.J., Regenerative medicine. Opportunities and challenges: A brief overview, J Royal Soc Interf7 (2010), S777–S781.
41.
NovoselE.C., KleinhansC. and KlugerP.J., Vascularization is the key challenge in tissue engineering, Adv Drug Del Rev63 (2011), 300–311.
42.
KimH.J., KimU.J., Vunjak-NovakovicG., MinB.H. and KaplanD.L., Influence of macroporous protein scaffolds on bone tissue engineering from bone marrow stem cells, Biomaterials26 (2005), 4442–4452.
43.
SafiniaL., MantalarisA. and BismarckA., Nondestructive technique for the characterization of the pore size distribution of soft porous constructs for tissue engineering, Langmuir22 (2006), 3235–3242.
44.
AlkhouliN., MansfieldJ., GreenE., BellJ., KnightB., LiversedgeN., ThamJ.C., WelbournR., ShoreA.C., KosK. and WinloveC.P., The mechanical properties of human adipose tissues and their relationships to the structure and composition of the extracellular matrix, Am J Physiol: Endocrinol Metab305 (1427), E1427–E1435.
45.
VukićevićR., NeffeA.T., GebauerT., FrankO., SchossigM. and LendleinA., Mechanical properties of architectured gelatin-based hydrogels on different hierarchical levels, MRS Adv1 (2016), 1995–2001.
46.
SartoriS., ChionoV., Tonda-TuroC., MattuC. and GianlucaC., Biomimetic polyurethanes in nano and regenerative medicine, J Mater Chem B2 (2014), 5128–5144.
