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Calcium nitrate hydrate

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Calcium nitrate hydrate is a chemical compound with the formula Ca(NO3)2·4H2O. It is a colorless, crystalline solid used as a source of calcium and nitrate ions in various applications. The core function of calcium nitrate hydrate is to provide calcium and nitrate ions for various chemical processes and applications.

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Lab products found in correlation

Calcium hydroxide, by Merck Group (8 mentions) Chondroitin 6 sulfate, by Merck Group (8 mentions)

8 protocols using calcium nitrate hydrate

1

Collagen-GAG Scaffold Fabrication and Crosslinking

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Col-GAG and MC-GAG scaffolds were prepared using lyophilization, as described previously (37 (link)–39 (link)). Briefly, microfibrillar, type I collagen (Collagen Matrix, Oakland, NJ) and chondroitin-6-sulfate (Sigma-Aldrich, St. Louis, MO) were combined in suspension in the absence and presence of calcium salts (calcium nitrate hydrate, Ca(NO3)2·4H2O; calcium hydroxide, Ca(OH)2; Sigma-Aldrich, St. Louis, MO) in an acetic acid (Col-GAG) or phosphoric acid (MC-GAG) solution. Using a constant cooling rate technique at a rate of 1°C/min, the solution was frozen from room temperature to −10°C using a freeze dryer (Genesis, VirTis). Following sublimation of the ice phase, scaffolds were sterilized via ethylene oxide and cut into 8-mm disks in diameter and 4 mm in height for culture.
Cross-linking of scaffolds was performed after rehydration in phosphate-buffered saline (PBS) for 4 hours using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDAC; Sigma-Aldrich) and N-hydroxysuccinimide (NHS; Sigma-Aldrich) at a molar ratio of 5:2:1 EDAC:NHS:COOH, where COOH represents the amount of collagen in the scaffold as we previously described (40 (link)). Scaffolds were washed with PBS to remove any of the residual chemical.
Ren X., Zhou Q., Foulad D., Tiffany A.S., Dewey M.J., Bischoff D., Miller T.A., Reid R.R., He T.C., Yamaguchi D.T., Harley B.A, & Lee J.C. (2019). Osteoprotegerin reduces osteoclast resorption activity without affecting osteogenesis on nanoparticulate mineralized collagen scaffolds. Science Advances, 5(6), eaaw4991.
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2

Fabrication of Collagen-GAG Scaffolds

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Collagen-GAG scaffolds were prepared using the lyophilization process described previously [18 (link),25 (link),26 (link)]. Briefly, a suspension of collagen and GAGs or collagen-glycosaminoglycan-calcium phosphate (CGCaP) were produced by combining microfibrillar, type I collagen (Collagen Matrix, Oakland, NJ) and chondroitin-6-sulfate (Sigma–Aldrich, St. Louis, MO) in a solution of 0.05 M acetic acid (pH 3.2) or with calcium salts (calcium nitrate hydrate: Ca(NO3)2·4H2O; calcium hydroxide: Ca(OH)2, Sigma–Aldrich) in a solution of phosphoric acid, respectively. The suspension was frozen using a constant cooling rate technique (1 °C/min) from room temperature to a final freezing temperature of −10 °C using a freeze dryer (Genesis, VirTis). The ice phase was sublimated under vacuum (<200 mTorr, 0 °C). Disks 5.8 mm in height and 8 mm in diameter were prepared using punch biopsies for cultures. Scaffold porosity was 85 ± 3% [27 (link)], pore size was 156 ± 6 μm [26 (link),27 (link)], and morphology consisted of isotropic pores with a transverse:longitudinal pore aspect ratio of 0.95 ± 0.01 [26 (link)] as we previously reported. All scaffolds were sterilized via ethylene oxide.
Ren X., Bischoff D., Weisgerber D.W., Lewis M.S., Tu V., Yamaguchi D.T., Miller T.A., Harley B.A, & Lee J.C. (2015). Osteogenesis on nanoparticulate mineralized collagen scaffolds via autogenous activation of the canonical BMP receptor signaling pathway. Biomaterials, 50, 107-114.
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3

Lyophilized Collagen-GAG Scaffold Fabrication

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Col-GAG and MC-GAG scaffolds were prepared using lyophilization as described previously (Harley, Leung, Silva, & Gibson, 2007 (link); Harley et al., 2010 (link); Weisgerber, Kelkhoff, Caliari, & Harley, 2013 (link)). Briefly, microfibrillar, type I collagen (Collagen Matrix, Oakland, NJ) and chondroitin-6-sulfate (Sigma-Aldrich, St. Louis, MO) were combined in suspension in the absence and presence of calcium salts (calcium nitrate hydrate: Ca(NO3)2·4H2O; calcium hydroxide: Ca(OH)2, Sigma-Aldrich, St. Louis, MO) in an acetic acid (Col-GAG) or phosphoric acid (MC-GAG) solution. Using a constant cooling rate technique at a rate of 1 °C/min, the solution was frozen from room temperature to −10 °C using a freeze dryer (Genesis, VirTis). Following sublimation of the ice phase, scaffolds were sterilized via ethylene oxide and cut into 8 mm disks for culture.
Crosslinking of scaffolds was performed after rehydration in phosphate buffered saline (PBS) for 4 hours using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC, Sigma-Aldrich) and N-hydroxysuccinimide (NHS, Sigma Aldrich) at a molar ratio of 5:2:1 EDC:NHS:COOH where COOH represents the amount of collagen in the scaffold as we previously described (Olde Damink et al., 1996 (link)). Scaffolds were washed with PBS to remove any of the residual chemical.
Ren X., Zhou Q., Foulad D., Dewey M.J., Bischoff D., Miller T.A., Yamaguchi D.T., Harley B.A, & Lee J.C. (2019). Nanoparticulate Mineralized Collagen Glycosaminoglycan Materials Directly and Indirectly Inhibit Osteoclastogenesis and Osteoclast Activation. Journal of tissue engineering and regenerative medicine, 13(5), 823-834.
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4

Fabrication of MC-GAG Scaffolds for Cell Culture

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MC-GAG scaffolds were prepared using the lyophilization process described previously [19 (link), 20 (link)]. Briefly, a suspension of collagen and GAGs were produced by combining microfibrillar, type I collagen (Collagen Matrix, Oakland, NJ) and chondroitin-6-sulfate (Sigma-Aldrich, St. Louis, MO) with calcium salts (calcium nitrate hydrate: Ca(NO3)2·4H2O; calcium hydroxide: Ca(OH)2, Sigma-Aldrich, St. Louis, MO) in a solution of phosphoric acid. The suspension was frozen using a constant cooling rate technique (1 °C/min) from room temperature to a final freezing temperature of −10 °C using a freeze dryer (Genesis, VirTis). Following sublimation of the ice phase, scaffolds were sterilized via ethylene oxide and cut into 6 or 8 mm diameter disks for culture. NX-MC scaffolds were rehydrated with phosphate buffered saline (PBS) overnight and used for cell culture. For stiffer MC scaffolds, scaffolds were rehydrated in PBS overnight with 1-ethyl-3-(3- dimethylaminopropyl) carbodiimide (EDAC, Sigma-Aldrich) and N-hydroxysuccinimide (NHS, Sigma Aldrich) at a molar ratio of 5:2:1 EDC:NHS:COOH where COOH represents the amount of collagen in the scaffold as we previously described [21 (link)].
Zhou Q., Ren X., Oberoi M.K., Bedar M., Caprini R.M., Dewey M.J., Kolliopoulos V., Yamaguchi D.T., Harley B.A, & Lee J.C. (2021). β-Catenin Limits Osteogenesis on Regenerative Materials in a Stiffness-Dependent Manner. Advanced healthcare materials, 10(23), e2101467.
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5

Lyophilized Collagen-GAG Scaffolds

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MC-GAG scaffolds were prepared using the lyophilization process described previously.[13 ,14 (link)] Briefly, a suspension of collagen and GAGs were produced by combining microfibrillar, type I collagen (Collagen Matrix, Oakland, NJ) and chondroitin-6-sulfate (Sigma-Aldrich, St. Louis, MO) with calcium salts (calcium nitrate hydrate: Ca(NO3)2·4H2O; calcium hydroxide: Ca(OH)2, Sigma-Aldrich, St. Louis, MO) in a solution of phosphoric acid. The suspension was frozen using a constant cooling rate technique (1 °C min−1) from room temperature to a final freezing temperature of −10 °C using a freeze dryer (Genesis, VirTis). Following sublimation of the ice phase, scaffolds were sterilized via ethylene oxide and cut into 8 mm diameter disks for culture. NX-MC scaffolds were rehydrated with phosphate buffered saline (PBS) overnight and used for cell culture.
Zhou Q., Lyu S., Bertrand A.A., Hu A.C., Chan C.H., Ren X., Dewey M.J., Tiffany A.S., Harley B.A, & Lee J.C. (2020). Stiffness of Nanoparticulate Mineralized Collagen Scaffolds Triggers Osteogenesis via Mechanotransduction and Canonical Wnt Signaling. Macromolecular bioscience, 21(3), e2000370.
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6

Preparation and Characterization of Collagen-GAG Scaffolds

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Col-GAG and MC-GAG scaffolds were prepared using lyophilization as we previously described [17, 18] . In brief, microfibrillar, type I collagen (Collagen Matrix, Oakland, NJ) and chondroitin-6-sulfate (Sigma-Aldrich, St. Louis, MO) were combined in suspension in an acetic acid solution to create Col-GAG scaffolds, versus in a phosphoric acid solution with calcium salts (calcium nitrate hydrate: Ca(NO 3 ) 2 •4H 2 O; calcium hydroxide: Ca(OH) 2 , Sigma-Aldrich) to create MC-GAG scaffolds. Using a freeze dryer (Genesis, VirTis, Gardiner, NY) and a constant cooling rate of 1 °C/min, the solution was frozen from room temperature to -10 °C. The scaffolds were then sublimated, sterilized with ethylene oxide, and stored at room temperature. For preparation for culture, the scaffolds were cut into 6-8 mm disks and rehydrated with ethanol, followed by phosphate buffered saline (PBS) overnight. The scaffolds were then crosslinked for four hours using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC, Sigma-Aldrich) and N-hydroxysuccinimide (NHS, Sigma-Aldrich) as previously described [19] . Scaffolds were washed with PBS to remove any residual chemicals.
Ren X., Zhou Q., Bedar M., Foulad D., Huang K.X., Dejam D., Dahan N.J., Kolliopoulos V., Harley B.A.C, & Lee J.C. (2023). Modulating Temporospatial Phosphate Equilibrium by Nanoparticulate Mineralized Collagen Materials Induces Osteogenesis via PiT-1 and PiT-2. Advanced healthcare materials, 12(17).
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7

Collagen-GAG Scaffold Fabrication and Sterilization

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MC-GAG scaffolds were prepared using the lyophilization process described previously (30) (31) (32) . Briefly, a suspension of collagen and GAGs were produced by combining microfibrillar, type I collagen (Collagen Matrix, Oakland, NJ) and chondroitin-6-sulfate (Sigma-Aldrich, St. Louis, MO) with calcium salts (calcium nitrate hydrate: Ca(NO 3 ) 2 •4H 2 O; calcium hydroxide: Ca(OH) 2 , Sigma-Aldrich, St. Louis, MO) in a solution of phosphoric acid. The suspension was frozen using a constant cooling rate technique (1 °C/min) from room temperature to a final freezing temperature of -10 °C using a freeze dryer (Genesis, VirTis). Following sublimation of the ice phase, scaffolds were sterilized via ethylene oxide and cut into 8 mm disks for culture. NX-MC scaffolds were rehydrated with phosphate buffered saline (PBS) overnight and used for cell culture.
Zhou Q., Lyu S., Bertrand A.A., Hu A.C., Chan C.H., Ren X., Dewey M.J., Tiffany A.S., Harley B.A., & Lee J.C. (2020). Stiffness of Nanoparticulate Mineralized Collagen Scaffolds Triggers Osteogenesis via Mechanotransduction and Canonical Wnt Signaling.
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8

Preparation and Characterization of MC-GAG Scaffolds

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MC-GAG scaffolds were prepared using the lyophilization process described previously (23) (24) (25) . Briefly, a suspension of collagen and GAGs were produced by combining microfibrillar, type I collagen (Collagen Matrix, Oakland, NJ) and chondroitin-6-sulfate (Sigma-Aldrich, St. Louis, MO) with calcium salts (calcium nitrate hydrate: Ca(NO3)2•4H2O; calcium hydroxide: Ca(OH)2, Sigma-Aldrich, St. Louis, MO) in a solution of phosphoric acid. The suspension was frozen using a constant cooling rate technique (1 °C/min) from room temperature to a final freezing temperature of -10 °C using a freeze dryer (Genesis, VirTis). Following sublimation of the ice phase, scaffolds were sterilized via ethylene oxide and cut into 6 or 8 mm diameter disks for culture. NX-MC scaffolds were rehydrated with phosphate buffered saline (PBS) overnight and used for cell culture. For stiffer MC scaffolds, scaffolds were rehydrated in PBS overnight with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC, Sigma-Aldrich) and Nhydroxysuccinimide (NHS, Sigma Aldrich) at a molar ratio of 5:2:1 EDC:NHS:COOH where COOH represents the amount of collagen in the scaffold as we previously described (26) .
Zhou Q., Ren X., Oberoi M.K., Caprini R.M., Dewey M.J., Kolliopoulos V., Yamaguchi D.T., Harley B.A., & Lee J.C. (2021). β-Catenin Limits Osteogenesis on Regenerative Materials in a Stiffness-Dependent Manner.
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