Nanocelluloses (NCs), with their remarkable characteristics, have proven to be one of the most promising “green” materials of our times and have received special attention from researchers in nanomaterials. A diversity of new functional materials with a wide range of biomedical applications has been designed based on the most desirable properties of NCs, such as biocompatibility, biodegradability, and their special physicochemical properties.
NCs Type |
Drug Delivery System |
Drug |
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Material |
Cellulose Source |
Toxicological Experiment Drug-Release Conditions |
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Material |
Material Cellulose Source |
Toxicological Experiment |
Drug Release |
Cells CellsLines Lines |
Toxicological ResultsMechanism |
Toxicological Ref. |
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Results and Possible Application |
Ref. |
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Cellulose Source |
Toxicological Experiment |
Cells Lines |
Toxicological Results |
Results Results and Possible Application |
Results and Possible Application Ref. |
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Ref. | ||||||||||||||||
CNC |
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CNC |
cCNC/SA double-membrane hydrogels |
CH, EGF |
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BNC | PBS, pH 7.4, 37 °C; |
t 90% = 3 days (CH); t90% = 4 to 8 days (EGF). |
Swelling/erosion |
[89] |
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TEMPO-oxidized CNC/CSos |
PrHy, IMI |
Breast cancer PBS, pH 7.4, room temp.; |
[50] |
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CNF |
CNC |
Cotton (Whatman 1 filter paper) t40% | ||||||||||||||
BNC scaffolds | = 12 min (PrHy); t 80% = 2 h (IMI). |
G. xylinus MTS assay; ATP assay. |
BEAS 2B | |||||||||||||
CNF | - |
hMDMs [ |
Bleached dissolving pulp Norway spruce (Picea bies) |
CCk-8 assay |
HUVECs, SMCs,90] |
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Cytotoxicity at 100 mg/mL; | No micronuclei induction after exposure to 2.5–100 mg/mL; |
MTT assay [3H]-thymidine uptake assayNo induction of proinflammatory cytokines in hMDMs. |
L929; Thymocytes Fibroblasts Toxicity impact on lungs or bone marrow |
PBMNCs |
CNFs were not cytotoxic; CNC has non-inflammatory and on-immunogenic properties. |
BC tubes have no toxic or side effects on vessel-related cells cultured on their surface; the surface of BC tubes was beneficial for cell attachment, proliferation, and ingrowth. Implantable biomaterials TE |
Vascular TE [135] |
[151] |
[166] |
CNC-HDQ complex |
HDQ |
Double crosslinking 3D-printed CNF hydrogels |
Skin TE dH2O, room temp., in the dark; t40% = 1 h; t |
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CNC CNC- carboxyl groups | ||||||||||||||||
CNF | 80% = 4 h. |
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Octenidine-loaded BNC |
[111] |
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Softwood cellulose pulp |
Pinus radiata pulp - |
MTS assay |
LDH assay MTT assay CaCO-2, HeLa, MDCK, J774 |
K. xylinus HEK NHDF [91] |
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ATP assay |
HaCaT CNC not exhibit any significant cytotoxicity; can exert stress on cells if they possess a high charge density; Charge-dependent decrease in mitochondrial activity (charge contents > 3.9 mmol/g). |
No toxic effect for keratinocytes and fibroblasts; Non-immunotoxic. Drug delivery |
[136] |
Pure BNC has no influence on HaCaT viability; OCT/BNC extracts exhibited time and concentration-dependent toxicity; cell-damaging effects were observed at extract conc >10% and longer incubation times (24 and 48 h). Wound dressings |
Active wound dressing [152] |
[167] |
CS/CNC nanocomposite hydrogels |
C |
CNF/PVA bilayer scaffold |
Skin TE SGF, pH 1.2, 37 |
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c-CNCs t-CNCs |
°C; |
Cotton Tunicate from Stuela clava [120 min: 65% (0.5% CS/CNC); 50% (2.5% CS/CNC). |
115] Ritger–Peppas model; n |
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CNF CNC | LDH assay | = 0.61–0.66; |
Wood pulp |
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BNC |
Sugar cane molasses TB assay Non-Fickian diffusion. |
A549 |
LDH activity MDM MDDC |
A549 HepG2/C3A [92] |
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The aspect ratio in combination with CNCs dose influences the uptake by the 3D co-culture system of the human epithelial airway barrier system. |
CNF caused a significant decrease in cell viability, at 72 h; Decrease in GSH levels after exposure to CNF. Toxicity impact on lungs |
BC is not cytotoxic (conc. < 170 μg/mL); BNC has a protective effect against CP-induced myelotoxicity and enotoxicity. CNC toxicity [137] |
Biomaterial TE [138] |
[168] |
QC/cCNC/β-GP nanocomposite hydrogels |
CNF/Gel/ApA DOX |
PBS, pH 7.4, 37 °C; t |
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U-NFC A-NFC C-NFC Bone TE | 90% = 4 days (0% cCNC); t |
[116] 90% = 7 days (1% cCNC); |
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CNC |
Wood pulp t90% = 17 days (2.5% cCNC). |
Never-dried bleached sulfite softwood dissolving pulp TB assay Swelling/erosion |
AB assay LDH assays A549 |
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Vaccarin- loaded BNC | Toxicity impacts on dermal, lung, and macrophage cells |
G. xylinus [ |
MTT assay HDF MRC-5 THP-1 [93] |
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CNC were nontoxic to A549 cells; | CNC induced a robust inflammatory response; |
CNC particles induced a more robust inflammatory response compared to NCF. |
No cytotoxicity for treated NFC; HDF and MRC-5 cells: the metabolic activity of the treated cells was comparable to that of the negative control; Comparable toxicity of CNC with CNF |
THP-1 cells: a higher metabolic activity of the NFC-treated; [153] |
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L929 |
BNC-Vac has lower toxicity and better biocompatibility than BNC; RGR for both BNC and BNC-Vac was above 74%. | 138] |
Wound dressing |
[169] |
Gel/CNC nanocomposite hydrogels |
CNC |
TPh |
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CNCgel CNC/PVA nanocomposites |
SGF, pH 1.2, 37 °C; |
Skin TE |
CNCdry 24 h: 90% (5% CNC); 85% (10% CNC); 60% (25% CNC). |
- [117] |
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Wood pulp |
LDH assay |
CNF |
[94] |
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Bleached Eucalyptus Globulus kraft pulp |
MH-S |
MTT assay Low conc. (1.5 and 5 μg/cm2) induce no cytotoxicity; A high dose of CNCdry induced a decrease in cell viability; CNC exposure further altered the secretion of cytokines. |
Toxicity impact on lungs |
[ |
A549 139] |
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Gentamycin-loaded BNC |
K. xylinus THP-1 |
NR assay Cytotoxic effect at the highest dose tested; Genotoxic effects in A549 cells in the co-cultures; No oxidative DNA damages. |
m-CNC/Alg hydrogels |
Ibu |
PBS, pH 7.4, 37 °C; t = 0–30 min; 45%–60% burst release; t = 30–330 min; sustained release. |
Fickian diffusion |
[95] |
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Gel/HA/CNC hydrogels |
Skin wound repair |
[118] |
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TE |
[ |
U2-OS 154] |
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No cytotoxicity on osteoblast culture after 24 h; | gentamycin released from G-BNC after 8 h (400 mg/L) and 16 h (600 mg/L) is enough to eliminate S. aureus and P. aeruginosa biofilms. |
Bone regeneration TE |
[170] |
CNC |
Wheat bran |
MTT assay |
CNF |
Curauá fibers (Ananas erectifolius L. B. Smith) |
Cytotoxicity assays ISO 10993-5 Caco-2 |
Vero |
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Curcumin- loaded BNC | Dose-dependent decrease in cell viability, but only with significant results above 1000 μg/mL; | The cell viability decreased significantly upon contact with CNC90 (88.09%) at 2000 μg/mL, although CNC30 (92.81%) and CNC60 (93.11%) did not significantly decrease the cell viability. |
CNF shows no cytotoxicity and suitable biocompatibility; The morphology and basic functions of the cells are not affected by the direct contact with the tested materials. |
K. xylinus Scaffold |
MTS assay Biocompatible nanocomposites |
HNDF |
The cytotoxic effect on the cells depended on the conc. of curcumin; at 0.5 mg/mL C, a strong cytotoxicity for BNC-C and BNC-DC180; BNC-DC300 suitable cytotoxicity, even at higher extract conc. TE [140] |
[ |
Wound dressing155] |
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[ | ] |
CNF |
PDA/TEMPO-CNF composite hydrogels |
TCH |
PBS; “On-off” drug release under NIR irradiation; 120 min: 60% (pH 5.0); 30% (pH 7.4); 15 h: 70% (pH 5.0); 55% (pH 7.4). |
Korsmeyer–Peppas model; Non-Fickian diffusion. |
Col/CNC/GMs |
Blood vessel |
CNF |
Softwood bleached kraft fiber |
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LDH | ||||||||||||||||
BNC-GTMAC BNC-GHDE | assay |
G. xylinus ] |
AB assay Caco-2, HT-29MTX Raji B |
Minimal or no cytotoxicity in a cellular model of the intestinal epithelium (for CNC-25 at 0.75% and 1.5% w/w, as well as for CNF-50 at 0.75% w/w). |
HaCaT |
No cytotoxicity; Suitable wound closure rates in the presence of the samples, with complete coverage of the scratched area after 5 days. Biocompatible material |
[156] |
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Wound dressing | [172] |
CNF/HPMC nanocomposites |
KT |
PBS, pH 7.4; 8 h: 95% (5% CNF), 62% (0.5% CNF), 56% (0.75% CNF), 37% (1% CNF). |
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PEG-grafted CNC nanocomposites |
Bone TE | Non-Fickian diffusion; n = 0.52–0.61. |
[ |
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CNF |
CNF/Alg hydrogels |
MH |
SGF, pH 1.2; SIF, pH 7.4, 37 °C; T40% = 90 min (CNF/Alg-50/50, SGF) t80% = 145 min (CNF/Alg-50/50, SIF). |
Fickian diffusion mechanism |
[98] |
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CNC/PVA hybrid hydrogels |
Soft TE |
[121] |
BNC |
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CNC/PAAm composite hydrogels |
BNC-SA hybrid hydrogels |
TE Ibu |
[122] PBS, pH 1.5, 7.0 and 11.8; 37 °C; 24 h: 90% (pH 11.8); 80% (pH 7.0); and 60% (pH 1.5); PBS, pH 7.4; 0.15 V, 0.3 V, 0.5 V; 24 h: 95% (0.5 V); 85% (0.3 V and 0.15 V); 80% (0 V). |
Korsmeyer–Peppas model; Non-Fickian diffusion; pH; n = 0.498–0.772; E-field; n = 0.700–0.491. |
[99] |
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TCH-loaded BNC composites |
TCH |
HEPES buffers, pH 7, 37 °C; 3 h: ~100% (free TCH); 90% (0.5% TCH); 60% (0.3% TCH); 20% (0.1% TCH); 10% (0.05% TCH); |
- |
[100] |
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OCT-loaded BNC |
OCT |
PBS, pH 7.4, 32 °C; 8 h: 82.7% ± 2.6% in first; 24 h 91.8% ± 2.0% after |
Ritger–Peppas model; n = 0.51–0.55; Non-Fickian diffusion. |
[101] |
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PI-loaded BNC |
PI |
PI buffer, 32 °C; t84% = 48 h. |
Ritger–Peppas model; n = 0.608–0.612 |
[102] |
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PHMB-loaded BNC |
PHMB |
PHMB buffer, 32 °C; t87% = 48 h. |
Ritger–Peppas model; n = 0.863–0.871 |
[102] |
Abbreviations: cCNC—Cationic cellulose nanocrystals; SA—Sodium alginate; CH—Ceftazidime hydrate; EGF—Epidermal growth factor human; PBS—Phosphate-buffered saline; CSos—Chitosan oligosaccharide; PrHy—Procaine hydrochloride; IMI—Imipramine hydrochloride; HDQ—Hydroquinone; C—Curcumin; CS—Chitosan; QC—Quaternized cellulose; β-GP—β-glycerophosphate; DOX—Doxorubicin; Gel—Gelatin; TPh—Theophylline; m-CNC—Magnetic cellulose nanocrystals; NIR—Near-infrared spectroscopy; PDA—Polydopamine; HPMC—Hydroxypropylmethyl cellulose; KT—Ketorolac tromethamine; Alg—Alginate; MH—Metformin hydrochloride; SGF—Simulated gastric fluid; SIF—Simulated intestinal fluid; HEPES—(4-(2-hydroxyethyl)-1-piperazine- ethanesulfonic acid); TCH—Tetracycline hydrochloride; AM—Acrylamide; Ibu—Ibupofren; OCT—Octenidine; PI—Povidone-iodine; PHMB—Polihexanide; dH2O—Distilled water.
NCs Type |
TE Systems |
Applications |
References |
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CNF |
CNF/CS nanocomposites |
Artificial skin |
[30] |
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CNF-based thixotropic gels | ||||||||||||||||||||||
U-CNF has an inflammatory response, which was suppressed when surface charges were introduced on the CNFs. | ||||||||||||||||||||||
K-CNC R-CNC |
Rubberwood fiber Kenaf-bast fiber |
MTT assay |
RAW 264.7 HaCaT |
Cytotoxicity of K-CNC and R-CNC is not significant up to 700 μg/mL; K-CNC and R-CNC induced the formation of ROS in RAW264.7 macrophages. |
Biocompatible nanocomposites |
[141] |
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CNC CNC-FL CNC-HM |
Cellulose pulp |
MTT assay |
ATCC PCS201012, A375 |
Banana peel bran No cytotoxicity in direct and indirect contact assays. |
Drug delivery |
[ |
MTT assay |
Caco-2 |
CNF conc. < 500 mg/mL are not cytotoxic to Caco-2 cells; Viability of Caco-2 decreased with increasing CNF conc. |
Biocompatible material] |
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BNC in nanocomposites | [ | 157] |
CNC in nanocomposites | |||||||||||||||||||
U-NFC A-NFC C-NFC P-NFC S-NFC |
Never-dried bleached sulfite softwood dissolving pulp |
Resazurin Assay |
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BNC/ALG bilayer composites |
G. xylinus Caco-2 |
ISO10993-5:2009 None of the NFCs inducing cytotoxic effects in the intestinal cells; |
hNCs hMNC The differences in physics-chemical properties of the studied NFCs were not reflected in the Caco-2 response in terms of metabolic activity and cell membrane integrity. |
Drug release in gastrointestinal tract (GIT) |
The composites were found to be noncytotoxic, with a cell viability of 98% and a uniform distribution of cells on the entire porous layer. [ |
Neocartilage TE 158] |
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[ | ] | Collagen/CNCs/ GMs scaffolds |
U-NFC C-NFC H-NFC MCC |
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BNC-COL-Ap compositesP-NFC S-NFC |
Never-dried bleached sulfite softwood dissolving pulp MTT assay |
HUVECs |
MTS Assay |
BEAS-2B |
G. xylinus |
MTT assay No cytotoxicity; Excellent biocompatibility. |
No cytotoxicity for the highest tested dose (500 μg/mL) for any of the NFCs; |
Osteoblastic cells Vascular TE |
None of the NFCs induced genotoxic effects; [143] |
All samples were able to increase intracellular formation of ROS. |
In vitro toxicity of NFCs |
The composites did not exhibit cytotoxicity effects. [ |
Bone regeneration TE 159] |
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[ | ] |
a-CNC/Gel hydrogels composite |
Breast cancer |
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CNC CNC-AEM CNC-AEMA |
c-CNF [123] |
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Softwood pulp |
ATP assay |
cys-CNF |
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ALG/BCN/COL composite Never-dried bleached sulfite softwood dissolving pulp | J774 A.1 PBMNC |
One cationic CNC induced secretion of proinflammatory cytokine IL-1b associated with increase mitochondrial-derived ROS and extracellular ATP levels. |
Drug and DNA delivery systems |
A. xylinum PB assay |
CCk-8 assay hDF |
MC3T3-E1 cys-CNF did not induce toxic effects on hDF when tested at a concentration up to 0.5 mg/mL, nor did the starting material c-NF cys-CNF presented a dual action in vitro: inhibition of metalloproteinase and radical scavenging activity. [144] |
hAMS Wound dressing |
MC3T3-E1 and hams cells were viable and proliferate well, after 2 and 5 days of incubation—suitable cytocompatibility. [ |
TE160] |
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[ | ] |
TEMPO-CNC reinforced PVA hydrogels |
CNF in nanocomposites |
Corneal implant |
[124] |
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PLA/CNCg-PEG nanocomposites |
BNC | |||||||||||||||||||||
BC-PHEMA |
BNC/Fibrin composites |
New blood vessel |
[78] |
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BNC-Gel/HAp nanocomposites |
Bone TE |
[125] |
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Southern |
pine |
Live/dead assays |
hMSCs |
Suitable biocompatibility; Nontoxic effect on hMSCs proliferation. |
Bone TE |
composites |
A. xylinum |
AB assay |
rMSCs |
BC-PHEMA composites are nontoxic and biocompatible; did not influence the morphology and proliferation of the rMSCs. [ |
Wound dressing 145] |
[176] |
CNF L-CNF |
Alg/BNC/Col composite |
TE |
[126] |
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3D BNC/PMS scaffolds |
TE; soft tissues regeneration |
[127] |
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DBC/Col-p |
TE; tissues regeneration |
[128] |
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BNC/PA/Gel/HAp | ||||||||||||||||||||||
TEMPO-CNC reinforced PVA hydrogels |
MCC |
AB assay |
HCE-2cells |
Nontoxicity; Excellent biocompatibility; The HCE-2 cells viability above the 70%. |
CNC L-CNC |
Dissolving pulp |
AB assay |
A549 THP-1 |
Cytotoxic and inflammatory responses were dependent on type, size, and hydrophobicity Low or inexistent toxicity of all CNMs in A549 cells | |||||||||||||
BC/COL composites |
G. xylinus |
Live/ Dead assayOphthalmic applications |
UCBMSCs Dose-dependent cytotoxic and inflammatory responses in THP-1 cells. |
No cytotoxicity; Provide advanced microenvironment for UCB-MSCs viability and in vitro proliferation;TE [146] |
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[ | ] |
PVA/CNC nanocomposites |
Sugarcane bagasse |
MTT assay |
L929 |
Noncytotoxic effect; Strong attachment and proliferation of human fibroblast skin cells on the scaffold. |
TE scaffolds |
[147] |
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GA-HA-CNC hydrogels |
MCC |
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Significantly elevated proteins and calcium deposition. | Bone regeneration TE |
[177] |
CNF /GEL/ApA |
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GEL/BNC Bleached birch pulp |
nanocomposite MTT assay |
A. xylinum |
MTT assay MSCs |
HEK293 CNFs and CNF-COOHs have no cytotoxicity; CNF-COOH-ApA cells expressed a low level of stress, visible through lower cell density and the cell inclusions. |
Bone TE |
BNG showed negligible cytotoxicity. [ |
Wound dressing 162] |
CCK-8 assay |
NIH-3T3 |
Cell viability, at 1, 4, and 7 days, higher than 70% limit; No foreign body response. |
Skin TE |
[148] |
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CS/Gel/NCC/CP nanocomposites |
Soft wood cellulose fibers |
MTT assay |
Fibroblast cell |
Lack of cytotoxicity after 3 days of increasing the cells’ viability. |
Wound healing |
[149] |
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CNC/PVA |
MCC |
AB assay |
HCE-2 |
Nontoxic and cytocompatible profile of the CNC-PVA hydrogel; Suitable biocompatibility toward HCE-2. |
Ophthalmic applications |
[150] |
Bone repair |
[129] |
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(BNC-Col)-Ap/OGP peptides |
Bone TE |
[130] |
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BNC-CNTs composites |
Bone regeneration |
[131] |
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BNC |
Ear cartilage TE |
[132] |
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BNC/Alg bilayer composite |
Neocartilage formation |
[133] |
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BNC/CS composites |
Cartilage tissue regeneration |
[134] |
Abbreviations: CS—Chitosan; PVA—Poly(vinyl) alcohol; Gel—Gelatin; ApA—Phosphonate; HA—Hyaluronic acid; Col—Collagen; GMs—Gelatin microspheres; PEG—Poly(ethylene glycol); PAAm—Polyacrylamide; a-CNC—Anionic CNC; HAp—Hydroxyapatite; Alg—Alginate; PMS—Paraffin microspheres; DBC—Dialdehyde bacterial cellulose; Col-p—Collagen peptide; PA—Procyanidin; Ap—Carbonate apatite; OGP—Osteogenic growth peptide; CNTs—Carbon nanotubes.
Abbreviations: BEAS 2B—Human bronchial epithelial cells; hMDMs—Monocyte-derived macrophages; Caco-2—Human colon carcinoma cells; HeLa—Human cervix; MDCK—Dog kidney; J774—Mouse macrophages; A549—Epithelial cells; MDM—Human blood monocyte-derived macrophages; MDDC—Dendritic cells; MH-S—Murine alveolar macrophages; RAW 264.7—Macrophages; ATCC PCS201012—Primary human fibroblasts; A375—Malignant melanoma cells; J774A.1—Mouse monocyte/macrophage; PBMNC—Peripheral blood mononuclear cells; hMSCs—Human mesenchymal stem cells; L929—Mouse fibroblast; NIH-3T3—Fibroblast; HCE-2—Human corneal epithelial cells; LDH assay—Lactate dehydrogenase cytotoxicity assay; MTT assay—(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay; TB assay—Trypan blue exclusion staining cell viability assay; MTS assay—CellTiter-Glo luminescent cell viability assay; MCC—Microcrystalline cellulose.
[ | |||||||||||
] | |||||||||||
NFC hydrogels crosslinked with Ca | |||||||||||
2+ | |||||||||||
Bleached sulfite softwood pulp |
AB assay |
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BNC-GEL nanocomposite | hDF |
- |
MTS assay Cell viability about 78% indicates no toxic effects. No inflammatory response of blood-derived mononuclear cells was observed in relation to the cytokines secretion. |
Wound healing |
MRC-5 |
The samples have no cytotoxicity, and the cells retained their morphology in direct contact with the membrane, The cells attaching to the GEL porous site, while not attaching to the GEL thin-coated BC side. |
Bone regeneration TE [163] |
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[ | ] |
TEMPO-CNF hydrogel |
Bleached birch kraft pulp |
MTT assay |
hDF |
Nontoxicity effect and great hDF cells viability. |
Wound healing |
[164] |
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NFC/QCRs nanocomposites |
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Chitosan-BNC |
K. xylinus |
MTT assay |
GM07492 |
No cytotoxicity for the BC group and BC-Chi-Cip group; Ciprofloxacin-loaded BC-Chi samples exhibited a significant but slight decrease in the metabolic activity of cells (moderate cytotoxicity). |
Wound dressing |
[180] |
Brown algae |
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GO/n-HAp/BNC/b-glucan biocomposite |
MTT assay |
- |
NR assay L929 |
Cells viability is higher than 80% (for 5 to 1000 μg/mL CNFs/QCRs), indicating that there is no cytotoxicity. |
MC3T3-E1 |
All samples had suitable potential for cell adhesion and proliferation with very low cytotoxicity The order of the cell viability: BgC-1.4 (93%) > BgC-1.3 (79.8%) > BgC-1.2 (71.4%) > BgC-1.1 (68.9%). Wound healing |
Bone regeneration TE [165] |
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[ | ] |
Abbreviations: HUVECs—Human umbilical vein endothelial cells; SMCs—Smooth muscle cells; HaCaT—Human keratinocytes cells; HepG2/C3A—Human C3A hepatoma cells; L929—Mouse skin fibroblast cells; U2-OS—Osteoblast cell; HNDF—Human neonatal dermal fibroblasts; MC3T3-E1—Mouse osteoblastic cells; rMSCs—Mouse mesenchymal stem cells; UCBMSCs—Human umbilical cord blood-derived mesenchymal stem cells; MRC-5—Normal lung tissues cells; GM07492—Human fibroblast cells; CCk-8 assay—Cholecystokinin-octopeptide proliferation assay; ATP assay—Adenosine triphosphate assay; LDH assay—Lactate dehydrogenase assay; NR assay—Neutral red assay; MTS assay—CellTiter 96® Aqueous Non-Radioactive Cell Proliferation assay; AB assay—Alamar blue assay; G. xylinus—Gluconacetobacter xylinus; K. xylinus—Komagataeibacter xylinus; A. xylinum—Acetobacter xylinum; BNC- GTMAC—BNC functionalized with glycidyl trime-thylammonium chloride (GTMAC); BNC-GHDE—BNC functionalized with glycidyl hexadecyl ether (GHDE).