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

Table 6.

Results of individual studies: Masson’s trichrome staining, ARS staining, OCN staining.

Studies Type of Study Biomaterials Bone Regeneration Measurement Results
[27] In vitro/in vivo (male rats)
  • a)

    CS (Qingdao Boyite Biomaterials Co. Ltd., Shandong, China) combined with CSn(pDNA-BMP2)-GP(*)

  • b)

    CS (Qingdao Boyite Biomaterials Co. Ltd., Shandong, China) combined with CSn-GP(*)

 Masson’s trichrome staining (width of new bone)
  • a)

    NB = 500 µm

  • b)

    NB = 300 µm
[33] In vivo (calvarial bone of mice)
  • a)

    CS combined with dicarboxylic acid + PDLCs

  • b)

    CS combined with dicarboxylic acid without PDLCs

 Masson’s trichrome staining Increase in the amount of collagen and bone matrix in CS/DA scaffold with and without PDLCs after 12 weeks
[38] In vivo (alveolar bone of male beagles)
  • a)

    CS anorganic bovine bone

  • b)

    CS anorganic bovine bone + hJBMMSCs

  • c)

    CS

  • d)

    CS + hJBMMSCs

  • e)

    anorganic bovine bone

  • f)

    Control group

 Masson’s trichrome staining CS + hJBMMSCs showed more dense and well-organized PDL than the other groups
[31] In vitro
  • a)

    CS combined with inorganic calcium phosphate and hyaluronic acid on which MSCs were cultured

  • b)

    control glass coverslip

 ARS staining of MSCs
  • a)

    Histological and immunohistochemical analysis of paraffin-embedded nodules evidenced the presence of mineralized matrix positive to red alizarin and cells positive to osteocalcin.

  • b)

    No major morphological changes and no nodules were observed
[36] In vivo (molar area of the mandibular body of rabbits)
  • a)

    PLGA nanoparticles (Shandong Institute of Medical Instruments) (100 mg PLGA dissolved in 10 mL acetone and slowly poured into 40 mL 2% polyvinyl alcohol) /CS nanoparticles (Tiengene Bio-Technique Co. Ltd., Guangzhou, China) (40 mg CS powder dissolved in 40 mL of 1% glacial acetic acid, then 10 mL of 0.1% TPP solution instilled into the CS solution)/Silver nanoparticles complex (Shanghai Chaowei Nanotechnology Co. Ltd. DMEM (HyClone)

 ARS staining of PDLCs More mineralized nodules were observed on nPLGA/nCS/nAG membrane than in negative control group
[39] In vivo (alveolar bone of male beagles)
  • a)

    200 mg CS (Sigma-Aldrich, St. Louis, MO, USA) combined with 56% w/v, 1 mL β-glycerophosphate (Merck, Darmstadt, Germany) thermosensitive hydrogel + 100 ng/mL BMP-7 (PeproTech, Rocky Hill, NJ, USA) and 0.5% ornidazole

  • b)

    200 mg CS (Sigma-Aldrich, St. Louis, MO, USA) combined with56% w/v, 1 mL β-glycerophosphate (Merck, Darmstadt, Germany) thermosensitive hydrogel + 100 ng/mL ornidazole

  • c)

    CS (Sigma-Aldrich, St. Louis, MO, USA) combined with 56% w/v, 1 mL β-glycerophosphate (Merck, Darmstadt, Germany thermosensitive hydrogel + 100 ng/mL BMP-7 (PeproTech, Rocky Hill, NJ, USA)

  • d)

    CS alone (Sigma-Aldrich, St. Louis, MO, USA)

 OCN staining for osteoblasts (number of OCN-positive cells)
  • a)

    OCN-positive cells = 45

  • b)

    OCN-positive cells = 22

  • c)

    OCN-positive cells = 43

  • d)

    OCN-positive cells = 19

AG = silver; ARS = ARS = alizarine red; BMP-7 = bone morphogenetic protein-7; CSn(pDNA-BMP2)-GP = chitosan nanoparticles loaded with bone morphogenetic protein-2 plasmid DNA into a chitosan-based hydrogel with α, β–glycerophosphate; DA = dicarboxylic acid; NB = new bone; hJBMMSCs = human jaw bone marrow-derived mesenchymal stem cells; MSCs = mesenchymal stem cells; PDLCs = periodontal ligament cells. (*) CS for nanoparticles: Hengtai Jinhu Crust Product Co. Ltd. (Shandong, China); αβ-GP: Sinopharm Chemical Reagent Co. Ltd.; Plasmid BMP2: Central Laboratory of the Affiliated Hospital of Qingdao University