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. 2020 Mar 25;10(4):605. doi: 10.3390/nano10040605

Table 4.

Results of individual studies: AlamarBlue and PrestoBlue assay.

Studies Type of Study Biomaterials Bone Regeneration Measurement Results
[22] In vitro
  • a)

    CS alone (Sigma-Aldrich, Germany)

  • b)

    1.1% (w/v) CS (Sigma-Aldrich, Germany) combined with hydroxyapatite

  • c)

    CS (Sigma-Aldrich, Germany) combined with hydroxyapatite and 1% (w/v) PCL (Sigma-Aldrich, Germany) solution in HFIP (Sigma-Aldrich, Germany)

  • d)

    CS (Sigma-Aldrich, Germany) combined with hydroxyapatite + 30 μL of BMP-6 solution including 100 ng of BMP-6 (recombinant human, R&D Systems, Minneapolis, MN, USA)

 PrestoBlue assay of MC3T3-E1 cells (absorbance values, 570 nm) at day 21
  • a)

    MC3T3-E1 = 0.72

  • b)

    MC3T3-E1 = 0.65

  • c)

    MC3T3-E1 = 0.67

  • d)

    MC3T3-E1 = 0.66
[25] In vitro/in vivo (calvarial bone defects of rabbits)
  • a)

    50 kDa CS

  • b)

    25 kDa CS

  • c)

    10 kDa CS

  • d)

    50 kDa CS + 1 mg/mL−1 osteoprotegerin (PeproTech, Rocky Hill, NJ, USA)

  • e)

    25 kDa CS + 1 mg/mL−1 osteoprotegerin (PeproTech, Rocky Hill, NJ, USA)

  • f)

    10 kDa CS + 1 mg/mL−1 osteoprotegerin (PeproTech, Rocky Hill, NJ, USA)

  • g)

    Control

 AlamarBlue assay (Sigma) of osteoblasts (metabolic viability %) seeded on the gels after 24 h
  • a)

    Viability = 140%

  • b)

    Viability = 120%

  • c)

    Viability = 120%

  • d)

    Viability = 150%

  • e)

    Viability = 140%

  • f)

    Viability = 150%

  • g)

    Viability = 110%
[29] In vitro
  • a)

    0.5g CS (Sigma Aldrich, St. Louis, MO, USA) + 1 g hyaluronic acid hydrogel (Sigma Aldrich, St. Louis, MO, USA) scaffold

  • b)

    1% (w/v) CS (Sigma Aldrich, St. Louis, MO, USA) hydrogel scaffold

  • c)

    Ha hydrogel (Sigma Aldrich, St. Louis, MO, USA) scaffold seeded with osteoblast and fibroblast cultures PBS solution without cells (negative control group)

 AlamarBlue assay (mitochondrial cell activity) The test showed a higher increase (20%) in cellular viability of both cellular groups compared with the control one. However, none of them were statistically different
[30] In vitro
  • a)

    0.7% (w/v) purified CS (Sigma-Aldrich) combined with 0.3% (w/v) bioactive-glass nanoparticles in a solution of 2 vol% acid acetic

  • b)

    CS alone (Sigma-Aldrich)

 AlamarBlue assay of PDLCs and hBMSCs Generally cell metabolic activity was higher in the CS/BG-NP
group compared with the CS group for both PDLCs and hBMSCs.
[32] In vitro/in vivo (8 weeks old rats)
  • a)

    Trilayered functionally-graded chitosan membrane (FGM) with bioactive glass gradient (50%, 25%, 0% wt.)

  • b)

    control group

 AlamarBlue assay of MC3T3-E1 cells (relative % AB reduction)
  • a)

    Relative % AB reduction = 150%

  • b)

    Relative % AB reduction = 90%

BMP-6 = bone morphogenetic protein-6; CS/BG-NP = chitosan/bioactive-glass nanoparticles; FGM = functionally-graded chitosan membrane; Ha = hyaluronic acid; hBMSCs = human bone marrow stromal cells; HFIP = 1,1,1,3,3,3-hexafluoro-2 propanol; PBS = phosphate buffer solution; PCL = polycaprolactone; PDLSCs = periodontal ligament stem cells;