Table 4.
Studies on CS-based scaffolds for cartilage tissue regeneration.
| Authors | Scaffold type | Study outline | Results | Conclusion |
|---|---|---|---|---|
| Chen et al. 2011 [80] | CS sponges (DA 15%; MW 400 kDa) +HA+GEL (ratio NA) whether or not activated by growth factors (BMP-2 and TGF) |
In vitro: CC and cell differentiation in MSC culture In vivo: implantation of sponges in osteochondral patellar defects of 4-month-old New Zealand white rabbits; HT and IHC evaluations Control group: DNA-free composite osteochondral graft Sample: n = 5 Control: n = 4 |
Growth and osteochondral differentiation in vitro were observed In vivo: greater osteochondral tissue neoformation with the scaffolds groups, with or without growth factors when compared to the control |
CS sponges + HA+GEL with TGF and BMP-2 promoted greater cell growth and bone and cartilage tissue regeneration (P < 0.05) |
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| Whu et al. 2013 [81] | CS (MW 65 kDa; DA 40%) +GEL (ratios 5 : 0, 4 : 1, 3 : 2, 1 : 1, 2 : 3, 1 : 4, or 0 : 5) in films and sponges, either crosslinked or not with carbodiimide | In vitro: CC in culture of chondrocytes with scaffolds In vivo: implantation of crosslinked scaffolds in cartilage defects in rabbits feet; HT and IHC analyses Control group: untreated defects Sample: n = 3 |
In vitro: greater cell proliferation and viability with crosslinked CS+GEL scaffolds In vivo: greater regeneration of cartilage with CS+GEL impregnated with chondrocytes after 1 month There was no comparison with pure CS or with absence of chondrocytes |
The authors conclude that carbodiimide crosslinked CS+GEL scaffold demonstrated potential for cartilage regeneration (P < 0.05) |
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| Zhang et al. 2013 [82] | Sponges of CS (MW 40 kDa; DA: NA) +PLGA (ratio 1 : 1), either with or without incorporation of SC | In vitro: CC in adipose-derived stem-cell culture, in chondrogenic medium In vivo: implantation of scaffolds, with or without SC, in articular defects in 4-month-old New Zealand rabbits knees; HT, IHC, and biomechanical assays (compressive modulus and cytonano-indentation) Control group: scaffold alone Sample: n = 5 |
In vitro: CS+PLGA favored chondrogenic adhesion, proliferation, and differentiation In vivo: CS+PLGA+SC promoted greater regeneration of the defects and maintenance of subchondral bone, after 12 weeks, and greater mechanical performance than scaffolds without SC |
CS+PLGA+SC scaffolds were capable of regenerating the full thickness of the cartilage defects in 12 weeks (P < 0.05) |
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| Deng et al. 2013 [83] | Sponges of CS (DD: NA; MW: NA) +SF (ratio 1 : 1); DA: NA; MW: NA incorporated or not with SC | In vitro: CC in BMMSC culture In vivo: filling of defects in the cartilage of 2-3- month-old New Zealand rabbit knees with scaffolds, with or without SC; HT and IHC analyses Control group: untreated group Sample: n = 6 |
In vitro: CS scaffolds promoted chondrogenic differentiation In vivo: the CS+SF+SC scaffold promoted almost complete repair of the defects and positive HT and IHC results; CS+SF scaffold demonstrated better results than the control, but not as good as in the group with SC |
CS+SF scaffold showed itself to be effective as SC carrier and capable of being used in the regeneration of cartilage tissue (P < 0.05) |
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| Wu et al. 2014 [84] | Sponges of pure CS (DA: NA; MW: NA) or combined with fibrin (ratio NA), whether or not incorporated with SC | In vitro: CC and chondrogenic differentiation in SC culture from synovial fluid In vivo: implantation in defects in 2-week-old nude mice TMJ disc; HT, IHC, and PCR analyses Control group: cell-free chitosan/fibrin scaffold Sample: n = 6 |
In vitro: CS + fibrin exhibited greater chondrogenic adhesion, proliferation, and differentiation In vivo: CS+SC and CS+fibrin+SC induced greater regeneration than acellular scaffolds at 4 weeks |
CS + fibrin scaffold with TMJ-derived stem cells demonstrated regenerative capacity for the treatment of TMJ disc perforations (P < 0.05) |
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| Cheng et al. 2014 [32] | Membranes of CS (DA ≤ 10%; MW: 200–500 kDa) +PLGA (ratio 75 : 25), whether or not impregnated with chondrocytes | In vitro: CC and chondrogenic differentiation inBMMSC culture In vivo: implantation in cartilage defects of 2-month-old New Zealand rabbit ears; macroscopic and HT analyses Control group: untreated defects Sample: n = 3 |
CS+PLGA + chondrocytes demonstrated complete and homogeneous regeneration after 18 weeks, with the formation of mature cartilage tissue; acellular scaffold and control group exhibited fibrosis | CS+PLGA impregnated with chondrocytes were capable of regenerating the cartilage tissue (P not reported) |
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| Ravanetti et al. 2015 [85] | CS+ raffinose (DA: NA; MW: NA; ratio: NA) | In vivo: implantation of scaffolds in osteochondral defects in the scapula of New Zealand white rabbits; macroscopic and HT analyses; negative control Control group: untreated defects Sample: n = 9 |
CS + raffinose did not promote regeneration of defects histologically or macroscopically and induced inflammation and formation of fibrous capsule, after 4 weeks | The authors conclude that CS + raffinose has limitations and that further studies are needed before application. P > 0.05 |
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| Ravindran et al. 2015 [86] | CS+COL (1 : 1), with or without SC and ECM (DA: NA; MW: NA) | In vitro: culture of MSC in osteogenic and chondrogenic medium In vivo: implantation in the subcutaneous tissue of mice; HT, IHC, and magnetic resonance analyses Control group: scaffolds without ECM Sample: not specified |
In vitro:ECM scaffolds induced chondrogenic differentiation In vivo: CS+COL+SC+ECM presented expression of chondrogenic differentiation markers after 2 weeks; with magnetic resonance, newly formed tissue similar to native cartilage was observed after 8 weeks |
The CS+COL+SC+ECM scaffold demonstrated efficiency in the regeneration of cartilage and bone tissue P < 0.01 |
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| Meng et al. 2015 [87] | CS hydrogel, either with or without DBM particles, E7 peptide (P7), and SC (DA = NA; MW = NA; ratio = NA) | In vitro: CC and chondrogenic differentiation in culture of BMSC; compression strength and elastic modulus tests In vivo: HT and IHC analyses of subcutaneous tissue of nude mice after 4 weeks Control group: pure CS scaffolds and composite scaffolds of DBM and CS; Sample: n = 5 |
In vitro: greater cell proliferation and differentiation with CS+DBM+P7 Preparation of DBM particles might influence the mechanical properties of scaffolds and cell proliferation In vivo: CS+DBM+P7+SC produced greater cartilage tissue formation than pure CS or with DBM; no negative control |
CS+DBM+P7 hydrogel combined with mesenchymal stem cells has potential for regeneration of cartilage tissue. P < 0.05 |
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| Zhang et al. 2015 [88] | CS sponges (MW 40 kDa; DA < 5%) +PLGA, with or without SC; ratio: NA; average viscosity | In vitro: CC and chondrogenic differentiation in culture of SC In vivo: implantation in cartilage defects in 4-month-old New Zealand rabbit knees; macroscopic, HT and IHC analyses; negative control not specified Control group: adherent ASC/scaffold complexes Sample: n = 5 |
In vitro: CS+PLGA+SC demonstrated chondrogenic differentiation In vivo: the scaffold promoted new formation of cartilage similar to hyaline, both histologically and via biomechanical evaluation, after 6 and 12 weeks |
CS+PLGA sponges incorporated with aggregated stem cells represents a promising technique in tissue regeneration P < 0.05 |
CC: cytocompatibility; COL: collagen; CS: chitosan; DA: degree of acetylation; DBM: Demineralized bone matrix; ECM: extracellular matrix; HT: histological; IHC: immunohistochemical; kDa: kilodaltons; MW: molecular weight; PCR: polymerase chain reaction; PLGA: polylactic-co-glycolic acid; SC/BMSC: stem cells/bone marrow stem cells; TMJ: temporomandibular joint.