Recent advances of natural biopolymeric culture scaffold: synthesis and modification

Figures & data

Table 1. Examples of commercial 3D culture scaffolds

Commercial 3D Culture Scaffold	Price	Manufacturer	Main composition of scaffold	References
Alvetex™ 3D Cell Growth Plates	$ 247.62 – $ 287.67 (pack of 2 plates)	Thermo Scientific	Synthetic polystyrene scaffold.	[8]
Cellusponge-Collagen	$ 1852.74 (pack of 48 plates)	Sigma	Plates coated with inert hydroxypropyl cellulose covered with collagen.	[9]
Nanofiber cell culture dish	$ 41.87 (per plate)	Sigma	Nanofiber cell culture dish, with aligned nanofibers.	[10]
PureCoat™ Amine 100 mm Dish	$ 144.60 (per case of 10)	Corning	A cell culture flask coated with fibronectin peptide.	[11]

Table 2. Recent reviews on culture scaffolds

Recent review papers	Authors and Year
Fabrication techniques of biomimetic scaffolds in three-dimensional cell culture: A review	[12]
Recent advances in ice templating: From biomimetic composites to cell culture scaffolds and tissue engineering	[21]
Colloidal systems toward 3D cell culture scaffolds	[18]
Scaffold-free 3-D cell sheet technique bridges the gap between 2-D cell culture and animal models	[19]
Scaffolds for 3D Cell Culture and Cellular Agriculture Applications Derived From Non-animal Sources	[22]
Graphene inks for the 3D printing of cell culture scaffolds and related molecular arrays	[13]
Bioreactor synergy with 3D scaffolds: New era for stem cells culture	[20]
Paper as a scaffold for cell cultures: Teaching an old material new tricks	[14]
Future of spermatogonial stem cell culture: Application of nanofiber scaffolds	[15]
Macroporous Hydrogel Scaffolds for Three-Dimensional Cell Culture and Tissue Engineering	[17]
Scaffolds for 3D in vitro culture of neural lineage cells	[16]

Table 3. Collagen culture scaffold

Crosslinkers	Mechanical properties	Porosity	Pore size	Type of cultured cells	Medium used	Maximum growth rate	References
1-ethyl-3-(3-dimethylamino-propyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS)	6.71 ± 3.2 kPA and 6.73 ± 1.7 kPA for collagen-only and collagen- chondroitin sulfate scaffolds	N/A	9.48 ± 4.7 µm	Primary porcine trabecular meshwork cells	DMEM with 10% FBS and 1% pen/strep	Fluorescent imaging with 90% or more of the top surface of the collagen scaffold was covered with cells	[25]
EDC	N/A	N/A	N/A	Multipotent human mesenchymal stem cells (hMSCs)	Dulbecco’s modified Eagle’s medium (DMEM), α-minimum essential medium (α-MEM)	2.4 × 10^6 cells/cm^3	[26]
Gradient crosslinked	N/A	N/A	1.6 × 10^8 pores/cm2	human small intestinal monolayers	expansion medium (EM)	Protein expression are more than 5 pmol/mg	[27]
N/A	Mechanical performance of the brain tissue: 6.9 kPa	N/A	N/A	Human neural stem cells, astrocytes, and microglia	N/A	cell viability of 84%	[30]
PFA, glutaraldehyde and osmium tetroxide 4 wt.%	Young’s modulus: 33 ± 12 kPa, Ultimate tensile Stress: 33 ± 6 kPa and Strain at failure: 105 ± 28%	N/A	25 to 95 µm	Normal Human Dermal Fibroblasts (NHDFs) and C2C12 murine myoblasts	low glucose Dulbecco’s modified Eagle’s medium (DMEM)	The cells show high proliferation rate under fluorescent microscopy	[31]
N/A	N/A	N/A	N/A	MLO-Y4 osteocytes	Minimum Essential Medium-α modification (α-MEM)	Fraction of dead cells of 0.0276 ± 0.0040	[33]
N/A	N/A	N/A	50 to 150 μm	human adipose-derived stem cells	DMEM	Quantification of differentiation rate of 75%	[29]
Glutaraldehyde	N/A	81 ± 3.9%	259.2 ± 45.2 µm	Hepatocyte (HepG2), liver cell cultivation	DMEM/high glucose	Cell proliferation with CCK-8 assay results of 2.7 at 450 nm absorbance, cell counting up to 5.8 × 10^5 cells per piece	[32]
N/A	Young’s Modulus: 26 kPa	N/A	N/A	Adipose-derived mesenchymal stem cells (ADSCs) and osteoblasts	Dulbecco’s modified Eagle’s medium	Cell proliferation assay by CCK-8 analysis for 21 days: 800%	[34]
EDC/NHS	Tensile Modulus: 31 ± 4.4 MPa	92.18 ± 2.05%	138.06 ± 59.40 µm	Human umbilical vein endothelial cells	Endothelial Cell Growth Medium 2	Percentage of EdU-positive cells 24 h after seeding: 27%	[28]
N/A	Max elevation: 0.05 mm	N/A	N/A	Human dermal fibroblast cell	Dulbecco’s Modified Eagle Medium	Cell viability (LDH assay): 53 U/L	[35]

Figure 1. Fluorescent photomicrographs of trabecular meshwork cells seeded on the surface of collagen only and collagen-chondroitin sulfate scaffolds cells scaffold. The cells were labeled with DAPI (blue) and glutaraldehyde fixation (green). 100,000 cells were seeded on each scaffold. In the z-stack images, the top surface of the scaffold is at the bottom of the image. Scale bars represent 50 µm [25].

Table 4. Chitosan Culture Scaffold

Crosslinkers	Mechanical properties	Porosity	Swelling degree	Type of cultured cells	Medium used	Maximum growth rate	References
N/A	Tensile strength: 1.33 ± 0.26 MPa	86.69 ± 1.97%	N/A	dental pulp stem cells	minimum essential medium Eagle	Cell number: 5 × 10^5 cells/scaffold	[36]
N/A	Young’s modulus of 27.7 kPa for 8% chitosan-hyaluronic acid	>90%	N/A	human glioblastoma cells (U-87 MG)	minimum essential medium	Cell number: 2.0 × 10^5 at day 9	[37]
Form of hydrogel	N/A	N/A	N/A	Bone marrow-derived mesenchymal stem cells	High glucose Dulbecco’s modi fied eagle’s medium	N/A	[38]
Form of hydrogel	Tensile strength: 4.1 MPa	N/A	50%	Mesenchymal stem cells	AmnioGrow, DMEM and KSFM	N/A	[40]
1-ethyl-3-(3-dimethylamino-propyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS)	N/A	N/A	N/A	Hepatocyte (Human liver cells)	Dulbecco’s modified Eagle’s medium (Hight Glucose)	Cell proliferation (CCK8 assay at 450 nm): 2.3	[39]
N/A	Stress at 50% compression: 12 kPa	68%	N/A	Osteoblast/Osteoclast (Bone cells)	alpha minimum essential medium	3.2 × 10^5 cells per scaffold	[41]

Figure 2. Gene expression and morphology of cells cultured on different substrates. (a) Relative expression displaying increased drug resistance and invasion specific genes in 3D scaffolds by U87-MG RFP cells after 12 days of culture (n = 3). Error bars represent standard error of mean. Scanning electron micrographs of GBM cells in (b) 2% Chitosan and hyaluronic acid (CHA), (c) 4% CHA, and (d) 8% CHA where tumor spheroid size increases with increasing stiffness. Histological staining (H&E) of tumor spheroids on (e) 2% CHA, (f) 4% CHA, and (g) 8% CHA scaffolds. Scale bars represent 100 μm (b-d) and 50 μm (e-g) [37].

Table 5. Silk fibroin culture scaffold

Crosslinkers	Mechanical properties	Porosity	Pore size	Swelling degree	Type of cultured cells	Medium used	Maximum growth rate	References
N/A	Interfacial strength: 57.3 ± 0.9 kPa	N/A	N/A	N/A	osteoblast and macrophage	Dulbecco’s modified Eagle’s medium	Bone formation: 70%	[42]
N/A	N/A	71 ± 2%	130 ± 23 μm	95.8 ± 1%	apical papilla stem cells	Dulbecco’s modified Eagle’s medium	DNA concentration: 120 ng/µl	[44]
N/A	N/A	42.06 ± 4.83%	100 µm to approximately 200 µm	N/A	Osteoblast	Dulbecco’s modified Eagle’s medium	Cell adhesion rate: 95%	[49]
N/A	Young’s modulus: 0.5 ± 0.6 MPa	N/A	N/A	N/A	bone marrow mesenchymal stem cells	Dulbecco’s modified Eagle’s medium	Cell viability: OD405 value 19.289 to 35.174	[43]
N/A	N/A	N/A	N/A	N/A	mesenchymal stem cells	Dulbecco’s modified Eagle’s medium	DNA quantification: 0.6 sGAG/DNA	[45]
N/A	N/A	82.2 ± 1.3%	3 to 212 µm	255%	mesenchymal stem cells	Dulbecco’s modified Eagle’s medium	Cell density: 900 cells/mm^2	[46]
N/A	N/A	86 to 90%	100.93 ± 1.36 μm	N/A	Human mesenchymal stem cells	Dulbecco’s modified Eagle’s medium	Cell distribution: 550 ± 42 cell/mm^2	[47]
N/A	N/A	>96%	150–170 µm	N/A	Articular chondrocytes and adipose-derived human mesenchymal stem cells	Dulbecco’s modified Eagle’s medium	DNA content: 7500 ng/g of scaffold	[48]
PAMAM dendrimers (0.5% EDC and 1% NHS)	N/A	N/A	N/A	N/A	L929 fibroblast cells	McCoy’s 5 A medium	Cell viability: 110%	[4]
N/A	Stress: 680 kPa	88.8%	300 um	758.9%	Stem cells from human exfoliated deciduous teeth (SHED)	α-modified Eagle’s medium	Albumin secreted: 1.4 g/dL	[50]

Figure 3. Confocal microscopy images of chondrogenic differentiated hMSCs on (a and c) SF/CS-Gl-Ch scaffolds and (b and d) pellet culture, where red represents immunofluorescence of Col II in a-b and Acan in c-d. Green fluorescence represents β-actin stained by phalloidin and blue corresponds to nucleus stained by Hoechst 33258 [47].

Table 6. Gelatin culture scaffold

Crosslinkers	Mechanical properties	Porosity	Pore size	Type of cultured cells	Medium used	Maximum growth rate	References
Into hydrogel form	N/A	2.99%	4 to 6 µm	HeLa cells (immortalized human cervical cancer cells)	Dulbecco’s modified eagle’s medium (DMEM)	Cell viability of 95%	[51]
N/A	1.26 ± 0.16 MPa	close to 90%	102.7 ± 39.9 µm	Mesothelial Cells and Mesothelium Tissue	Dulbecco’s Modified Eagles Medium (DMEM) and fetal bovine serum (FBS)	DNA content of cells: 1.38 µg	[52]
Ethylene dichloride (EDC)/N-hydroxysuccinimide (NHS)	N/A	N/A	N/A	Cornea epithelial cells	Dulbecco’s Modified Eagles Medium	Cell number: 2.8 × 10^4 to 3.8 × 10^4 cells	[53]
EDC/NHS	4 MPa	N/A	N/A	Endothelial cells and smooth muscle cells	Fetal calf serum (FCS) and Minimum Essential Medium	Increased significantly in HE staining results	[54]

Figure 4. 3D culture in porous structures of gelatin-methacrylate hydrogel scaffolds. (a) Representative scanning electron micrographs showing porous structures of gelatin-methacrylate hydrogel scaffolds fabricated with no porogen (left) and 15% (w/v) of gelatin porogen (right). (b) Bar graph presenting porosity measured in the gelatin-methacrylate scaffolds shown in (A). Error bars represent standard deviation. (c) Optical micrograph showing a HeLa cell-seeded gelatin-methacrylate scaffold at seeding density of 5 × 10⁶ cell/ml. White dotted squares (center and border) indicate regions of interest for estimation of cell viability as presented in (d) and (e). (D) Fluorescence micrographs showing live (green) and dead (red) cells at the center (top) and border (bottom) in gelatin-methacrylate scaffolds fabricated with no porogen (left) and 15% (w/v) of gelatin porogen (right). (E) Bar graph displaying cell viability at the center (white) and border (gray) in gelatin-methacrylate scaffolds fabricated with no porogen and 15% (w/v) of gelatin porogen [51].

Table 7. Cellulosic culture scaffold

Type of cellulose	Mechanical properties	Porosity	Pore size	Type of cultured cells	Medium used	Maximum growth rate	References
Bacterial cellulose	Tensile strength: 157.90 ± 23 MPa	N/A	N/A	Epidermal cells cultivation	Dulbecco’s modified Eagle’s medium	Cell area: 83.3 µm^2	[57]
Cellulose acetate	Elongation at break: 247.00 ± 47.27%	91.83 ± 14.62%	N/A	Bacteria cells S. aureus, S. epidermidis, and E. coli	Nutrient agar	Reduced bacteria inhibition	[58]
Bacterial cellulose	N/A	N/A	N/A	Human dermal fibroblast cultivation	Dulbecco’s modified Eagle’s medium	Antibacterial rates: 96.6 ± 0.5% against S. aureus and 90.6 ± 1.6% against E. coli.	[60]
Bacterial cellulose	N/A	N/A	N/A	U251 Human glioblastoma cancer cell line	Dulbecco’s modified Eagle’s medium	Cell viability: 110%	[59]

Figure 5. Schematic diagram of BC based scaffolds [59].

Table 8. Agarose culture scaffolds

Crosslinkers	Mechanical properties	Porosity	Pore size	Type of cultured cells	Medium used	Maximum growth rate	References
N/A	N/A	N/A	N/A	neuron cells for neuroregeneration	Dulbecco’s modified Eagle’s medium	Cell viability: 1.1	[61]
N/A	N/A	N/A	N/A	Corneal epithelial cells	Dulbecco modifid Eagle medium	Wound healing surface area: 25 mm^2	[62]
N/A	N/A	N/A	N/A	Human dermal fibroblasts	Dulbecco’s modified Eagle’s medium	Cell growth rate:182% at day 7	[63]

Scientific TF. Products. 2021.

 Google Scholar

Merck S-A. Cellusponge-Collagen. 2021.

 Google Scholar

Merck S-A Nanofiber cell culture dish, with random oriented nanofibers. 2021.

 Google Scholar

Corning LS Corning® PureCoat™ Amine 100 mm Dish, 10/Pack, 40/Case. 2021.

 Google Scholar

Badekila AK, Kini S, Jaiswal AK. Fabrication techniques of biomimetic scaffolds in three-dimensional cell culture: a review. J Cell Physiol. 2021;236(2):741–762.

 PubMed Web of Science ®Google Scholar

Qin K, Parisi C, Fernandes FM. Recent advances in ice templating: from biomimetic composites to cell culture scaffolds and tissue engineering. J Mat Chem B. 2021;9(4):889–907.

 PubMed Web of Science ®Google Scholar

Vila-Parrondo C, García-Astrain C, Liz-Marzán LM. Colloidal systems toward 3D cell culture scaffolds. Adv Colloid Interface Sci. 2020 283 ;102237.

 PubMedGoogle Scholar

Alghuwainem A, Alshareeda AT, Alsowayan B. Scaffold-free 3-D cell sheet technique bridges the gap between 2-D cell culture and animal models. Int J Mol Sci. 2019;20:4926.

 PubMedGoogle Scholar

Campuzano S, Pelling AE. Scaffolds for 3D Cell Culture and Cellular Agriculture Applications Derived From Non-animal Sources. Front Sustainable Food Syst. 2019;3:38.

 Google Scholar

Vlăsceanu GM, Iovu H, Ioniţă M. Graphene inks for the 3D printing of cell culture scaffolds and related molecular arrays. Compos Part B Eng. 2019;162:712–723.

 Web of Science ®Google Scholar

Yi T, Huang S, Liu G, et al. Bioreactor synergy with 3D scaffolds: new era for stem cells culture. ACS Appl Bio Mater. 2018;1(2):193–209.

 PubMedGoogle Scholar

Wu X, Suvarnapathaki S, Walsh K, et al. Paper as a scaffold for cell cultures: teaching an old material new tricks. MRS Commun. 2018;8(1):1–14.

 Web of Science ®Google Scholar

Shams A, Eslahi N, Movahedin M, et al. Future of spermatogonial stem cell culture: application of nanofiber scaffolds. Curr Stem Cell Res Ther. 2017;12(7):544–553

 PubMed Web of Science ®Google Scholar

Fan C, Wang DA. Macroporous Hydrogel Scaffolds for Three-Dimensional Cell Culture and Tissue Engineering. Tissue Eng Part B: Rev. 2017;23(5):451–461

 PubMed Web of Science ®Google Scholar

Murphy AR, Laslett A, O’Brien CM, et al. Scaffolds for 3D in vitro culture of neural lineage cells. Acta Biomater. 2017;54:1–20.

 PubMed Web of Science ®Google Scholar

Osmond M, Bernier SM, Pantcheva MB, et al. Collagen and collagen-chondroitin sulfate scaffolds with uniaxially aligned pores for the biomimetic, three dimensional culture of trabecular meshwork cells. Biotechnol Bioeng. 2017;114(4):915–923.

 PubMed Web of Science ®Google Scholar

Pustlauk W, Paul B, Brueggemeier S, et al. Modulation of chondrogenic differentiation of human mesenchymal stem cells in jellyfish collagen scaffolds by cell density and culture medium. J Tissue Eng Regen Med. 2017;11(6):1710–1722.

 PubMed Web of Science ®Google Scholar

Speer JE, Gunasekara DB, Wang Y, et al. Molecular transport through primary human small intestinal monolayers by culture on a collagen scaffold with a gradient of chemical cross-linking. J Biol Eng. 2019;13:13.

 PubMed Web of Science ®Google Scholar

Van Drunen R, Jimenez-Vergara AC, Tsai EH, et al. Collagen Based Multicomponent Interpenetrating Networks as Promising Scaffolds for 3D Culture of Human Neural Stem Cells, Human Astrocytes, and Human Microglia. ACS Appl Bio Mater. 2019;2(3):975–980.

 PubMedGoogle Scholar

Rieu C, Parisi C, Mosser G, et al. Topotactic Fibrillogenesis of Freeze-Cast Microridged Collagen Scaffolds for 3D Cell Culture. ACS Appl Mater Interfaces. 2019;11(16):14672–14683.

 PubMed Web of Science ®Google Scholar

Fournier R, Harrison RE. Methods for studying MLO-Y4 osteocytes in collagen-hydroxyapatite scaffolds in the rotary cell culture system. Connect Tissue Res. 2020;62(4):436–453.

 PubMed Web of Science ®Google Scholar

Li M, Ma J, Gao Y, et al. Epithelial differentiation of human adipose-derived stem cells (hASCs) undergoing three-dimensional (3D) cultivation with collagen sponge scaffold (CSS) via an indirect co-culture strategy. Stem Cell Res Ther. 2020;11(1):11.

 PubMed Web of Science ®Google Scholar

Meng D, Lei X, Li Y, et al. Three dimensional polyvinyl alcohol scaffolds modified with collagen for HepG2 cell culture. J Biomater Appl. 2020;35(4–5):459–470.

 PubMed Web of Science ®Google Scholar

Kim H, Han SH, Kook YM, et al. A novel 3D indirect co-culture system based on a collagen hydrogel scaffold for enhancing the osteogenesis of stem cells. J Mat Chem B. 2020;8(41):9481–9491.

 PubMed Web of Science ®Google Scholar

Malcor JD, Hunter EJ, Davidenko N, et al. Collagen scaffolds functionalized with triple-helical peptides support 3D HUVEC culture. Regen Biomater. 2021;7(5):471–482.

 Web of Science ®Google Scholar

Imashiro C, Yamasaki K, Tanaka RI, et al. Perfusable system using porous collagen gel scaffold actively provides fresh culture media to a cultured 3d tissue. Int J Mol Sci. 2021;22:6780.

 PubMedGoogle Scholar

Westin CB, Trinca RB, Zuliani C, et al. Differentiation of dental pulp stem cells into chondrocytes upon culture on porous chitosan-xanthan scaffolds in the presence of kartogenin. Mater Sci Eng C. 2017;80:594–602.

 PubMed Web of Science ®Google Scholar

Erickson AE, Lan Levengood SK, Sun J, et al. Fabrication and Characterization of Chitosan–Hyaluronic Acid Scaffolds with Varying Stiffness for Glioblastoma Cell Culture. Adv Healthc Mater. 2018;7(15):1800295.

 Web of Science ®Google Scholar

Qu C, Bao Z, Zhang X, et al. A thermosensitive RGD-modified hydroxybutyl chitosan hydrogel as a 3D scaffold for BMSCs culture on keloid treatment. Int J Biol Macromol. 2019;125:78–86.

 PubMed Web of Science ®Google Scholar

Radwan-Pragłowska J, Piątkowski M, Kitala D, et al. Microwave-assisted synthesis and characterization of bioactive chitosan scaffolds doped with Au nanoparticles for mesenchymal stem cells culture. Int J Polym Mater Polym Biomater. 2019;68(7):351–359.

 Web of Science ®Google Scholar

Zhao C, Li Y, Peng G, et al. Decellularized liver matrix-modified chitosan fibrous scaffold as a substrate for C3A hepatocyte culture. J Biomater Sci Poly Ed. 2020;31(8):1041–1056.

 PubMed Web of Science ®Google Scholar

Kruppke B, Heinemann C, Farack J, et al. Hemocyanin modification of chitosan scaffolds with calcium phosphate phases increase the osteoblast/osteoclast activity ratio—A co-culture study. Molecules. 2020; 25:4580.

 PubMedGoogle Scholar

Bhattacharjee P, Maiti TK, Bhattacharya D, et al. Effect of different mineralization processes on in vitro and in vivo bone regeneration and osteoblast-macrophage cross-talk in co-culture system using dual growth factor mediated non-mulberry silk fibroin grafted poly (Є-caprolactone) nanofibrous scaffold. Colloids Surf B Biointerfaces. 2017;156:270–281.

 PubMed Web of Science ®Google Scholar

Amirikia M, Shariatzadeh SMA, Jorsaraei SGA, et al. Impact of pre-incubation time of silk fibroin scaffolds in culture medium on cell proliferation and attachment. Tissue Cell. 2017;49(6):657–663.

 PubMed Web of Science ®Google Scholar

Li J, Wang Q, Gu Y, et al. Production of composite scaffold containing silk fibroin, chitosan, and gelatin for 3D cell culture and bone tissue regeneration. Med Sci Monit. 2017;23:5311–5320.

 PubMed Web of Science ®Google Scholar

Nalvuran H, Elçin AE, Elçin YM. Nanofibrous silk fibroin/reduced graphene oxide scaffolds for tissue engineering and cell culture applications. Int J Biol Macromol. 2018;114:77–84.

 PubMed Web of Science ®Google Scholar

Agrawal P, Pramanik K, Biswas A, et al. In vitro cartilage construct generation from silk fibroin- chitosan porous scaffold and umbilical cord blood derived human mesenchymal stem cells in dynamic culture condition. J Biomed Mater Res - Part A. 2018;106(2):397–407

 PubMed Web of Science ®Google Scholar

Agrawal P, Pramanik K, Vishwanath V, et al. Enhanced chondrogenesis of mesenchymal stem cells over silk fibroin/chitosan-chondroitin sulfate three dimensional scaffold in dynamic culture condition. J Biomed Mater Res B Appl Biomater. 2018;106(7):2576–2587.

 PubMed Web of Science ®Google Scholar

Agrawal P, Pramanik K. Enhanced chondrogenic differentiation of human mesenchymal stem cells in silk fibroin/chitosan/glycosaminoglycan scaffolds under dynamic culture condition. Differentiation. 2019;110:36–48.

 PubMed Web of Science ®Google Scholar

Bhardwaj N, Singh YP, Mandal BB. Silk Fibroin Scaffold-Based 3D Co-Culture Model for Modulation of Chondrogenesis without Hypertrophy via Reciprocal Cross-talk and Paracrine Signaling. ACS Biomater Sci Eng. 2019;5(10):5240–5254.

 PubMed Web of Science ®Google Scholar

Pourabadeh A, Mirjalili M, Shahvazian M. Modification of silk fibroin nanofibers scaffold by PAMAM dendrimer for cell culture. J Exp Nanosci. 2020;15(1):297–306.

 Web of Science ®Google Scholar

Huang TY, Wang GS, Ko CS, et al. A study of the differentiation of stem cells from human exfoliated deciduous teeth on 3D silk fibroin scaffolds using static and dynamic culture paradigms. Mater Sci Eng C. 2020;109:110563.

 PubMedGoogle Scholar

Shim K, Kim SH, Lee D, et al. Fabrication of micrometer-scale porous gelatin scaffolds for 3D cell culture. J Ind Eng Chem. 2017;50:183–189.

 Web of Science ®Google Scholar

Kao HH, Kuo CY, Chen KS, et al. Preparation of gelatin and gelatin/hyaluronic acid cryogel scaffolds for the 3D culture of mesothelial cells and mesothelium tissue regeneration. Int J Mol Sci. 2019;20 :4527.

 PubMedGoogle Scholar

Shahin A, Ramazani SAA, Mehraji S, et al. Synthesis and characterization of a chitosan/gelatin transparent film crosslinked with a combination of EDC/NHS for corneal epithelial cell culture scaffold with potential application in cornea implantation. Int J Polym Mater Polym Biomater. 2020:1865349. https://doi.org/10.1080/00914037.2020.1865349. .

 Google Scholar

Zhang Y, Jiao Y, Wang C, et al. Design and characterization of small-diameter tissue-engineered blood vessels constructed by electrospun polyurethane-core and gelatin-shell coaxial fiber. Bioengineered. 2021;12(1):5769–5788.

 PubMed Web of Science ®Google Scholar

Rao SN, Tsujita Y, Kondo T. Surface modification of oriented polysaccharide scaffolds using biotic nanofibers for epidermal cell culture. Cellulose. 2019;26(13–14):7971–7981.

 Web of Science ®Google Scholar

Felgueiras HP, Teixeira MA, Tavares TD, et al. Antimicrobial action and clotting time of thin, hydrated poly(vinyl alcohol)/cellulose acetate films functionalized with LL37 for prospective wound-healing applications. J Appl Polym Sci. 2020;137:48626.

 Google Scholar

He W, Zhang Z, Zheng Y, et al. Preparation of aminoalkyl-grafted bacterial cellulose membranes with improved antimicrobial properties for biomedical applications. J Biomed Mater Res Part A. 2020;108(5):1086–1098.

 PubMed Web of Science ®Google Scholar

Unal S, Arslan S, Yilmaz BK, et al. Production and characterization of bacterial cellulose scaffold and its modification with hyaluronic acid and gelatin for glioblastoma cell culture. Cellulose. 2021;28(1):117–132.

 Web of Science ®Google Scholar

Atoufi Z, Zarrintaj P, Motlagh GH, et al. A novel bio electro active alginate-aniline tetramer/ agarose scaffold for tissue engineering: synthesis, characterization, drug release and cell culture study. J Biomater Sci Polym Ed. 2017;28(15):1617–1638.

 PubMed Web of Science ®Google Scholar

Berkowski WM, Gibson DJ, Craft SL, et al. Development and assessment of a novel ex vivo corneal culture technique involving an agarose-based dome scaffold for use as a model of in vivo corneal wound healing in dogs and rabbits. Am J Vet Res. 2020;81(1):47–57.

 PubMed Web of Science ®Google Scholar

Yamada Y, Yoshida C, Hamada K, et al. Development of Three-Dimensional Cell Culture Scaffolds Using Laminin Peptide-Conjugated Agarose Microgels. Biomacromolecules. 2020;21(9):3765–3771.

 PubMed Web of Science ®Google Scholar