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2 protocols using metanol

1

Chitosan-Silica Hybrid Biomaterial Synthesis

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Tetraethylortosilicate (TEOS, 99%) and Chloride acid (HCl) (37%) were obtained from Alfa Aesar (Haverhill, MA, USA). Chitosan (CS; 50,000–190,000 Da; 75%–85% deacetylation degree) and 3-glycidoxypropyltrimethoxysilane (GPTMS, >98%) were purchased for Sigma Aldrich (St. Louis, MO, USA). Absolute ethanol (99.5%) was purchased from Panreac (Barcelona, Spain). Acid acetic (Reagent Grade) was purchased for Scharlau (Barcelona, Spain). HOB® human osteoblasts, foetal calf serum and Osteoblast Growing Medium (Promocell, Heidelberg, Germany). Paraformaldehyde, PBS, Triton x-100, bovine serum albumin, Metanol, rhodamine phalloidin and monoclonal anti-vinculin FITC conjugate were all purchased from Sigma, (St. Louis, MI, USA), Vectashield® (Vector, Burlingame, CA, USA).
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2

Chitosan-Silicate Scaffold for Osteoblast Culture

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Chitosan (CS; 50,000–190,000 Da; 75–85% deacetylation degree) was supplied by Sigma Aldrich (St. Louis, MI, USA). Tetraethylortosilicate (TEOS, 99%) and hydrochloric acid (37%, Pharma grade) were purchased from Alfa Aesar (Haverhill, MA, USA). Tri-calciumphosphate (TCP, pure, pharma grade) and absolute ethanol (99.5%) were obtained from Panreac (Barcelona, Spain), HOB® human osteoblasts, fetal calf serum, and Osteoblast Growing Medium (Promocell, Heidelberg, Germany) Paraformaldehyde, PBS, Triton x-100, bovine serum albumin, Metanol, rhodamine phalloidin, and monoclonal anti-vinculin FITC conjugate were all purchased from Sigma Aldrich, (St. Louis, MI, USA) and Hard Set Vectashield with DAPI® (Vector, Burlingame, CA, USA).
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