Figures & data
Table 1 Physicochemical properties of the calcium carbonates examined
Figure 1 XRD patterns and SEM images of B-Cal (a) and N-Cal (b) as produced (A) and after being incubated in simulated gastric (B) and intestinal fluids (C) and plasma (D).
Notes: (hkl) Miller indexing in XRD patterns (A) are typical patterns of the calcite phase (JCPDS No 47-1743); Asterisks in XRD patterns stand for the evolution of dicalcium phosphate dihydrate (JCPDS No 72-0714); SEM images were obtained after incubation for 1 hour in simulated gastric fluid and 4 hours in simulated intestinal fluid or plasma condition.
Abbreviations: arb, arbitrary; B-Cal, bulk calcium carbonates; hkl, Miller indexing; JCPDS, Joint Committee on Powder Diffraction Standards; N-Cal, nano calcium carbonates; SEM, scanning electron microscopy; XRD, X-ray diffraction; h, hours.
![Figure 1 XRD patterns and SEM images of B-Cal (a) and N-Cal (b) as produced (A) and after being incubated in simulated gastric (B) and intestinal fluids (C) and plasma (D).Notes: (hkl) Miller indexing in XRD patterns (A) are typical patterns of the calcite phase (JCPDS No 47-1743); Asterisks in XRD patterns stand for the evolution of dicalcium phosphate dihydrate (JCPDS No 72-0714); SEM images were obtained after incubation for 1 hour in simulated gastric fluid and 4 hours in simulated intestinal fluid or plasma condition.Abbreviations: arb, arbitrary; B-Cal, bulk calcium carbonates; hkl, Miller indexing; JCPDS, Joint Committee on Powder Diffraction Standards; N-Cal, nano calcium carbonates; SEM, scanning electron microscopy; XRD, X-ray diffraction; h, hours.](/cms/asset/e610e4f1-f773-4623-940b-ed0ee1ecf431/dijn_a_79403_f0001_b.jpg)
Table 2 In vitro and ex vivo dissolution and protein fluorescence quenching ratio of calcium carbonate materials
Table 3 List of the most abundant plasma proteins bound on B-Cal and N-Cal as determined by LC–MS
Figure 2 Plasma protein binding profiles of calcium carbonates separated by one-dimensional (A), and two-dimensional (B), gel electrophoresis.
Abbreviations: B-Cal, bulk calcium carbonates; N-Cal, nano calcium carbonates.
![Figure 2 Plasma protein binding profiles of calcium carbonates separated by one-dimensional (A), and two-dimensional (B), gel electrophoresis.Abbreviations: B-Cal, bulk calcium carbonates; N-Cal, nano calcium carbonates.](/cms/asset/8d66b265-ac6d-4413-9cb5-27eaefe6b983/dijn_a_79403_f0002_c.jpg)
Table 4 Biokinetic parameters and absorption of calcium carbonates after oral administration to rats
Figure 3 Plasma concentration-time curves (A–D) and tissue distribution (E, F) of calcium carbonates after a single (A, B), or 14 days of daily oral treatment (C–F) to rats.
Notes: Biokinetic data are presented as increase in calcium levels after subtracting the basal plasma calcium levels observed in untreated controls; (A) Biokinetics after administering a single dose to male rats; (B) Biokinetics after administering a single dose to female rats; (C) Biokinetics after 14-days of daily oral treatment in male rats; (D) Biokinetics after 14-days of daily oral treatment in female rats; (E) Tissue distribution in male rats; (F) Tissue distribution in female rats. (a) and (b) is used when there is no significant difference between two groups (control and N-Cal/N-Cal and B-Cal), but significant difference is found when three groups (control, N-Cal, B-Cal) are compared.
Abbreviations: B-Cal, bulk calcium carbonates; N-Cal, nano calcium carbonates.
![Figure 3 Plasma concentration-time curves (A–D) and tissue distribution (E, F) of calcium carbonates after a single (A, B), or 14 days of daily oral treatment (C–F) to rats.Notes: Biokinetic data are presented as increase in calcium levels after subtracting the basal plasma calcium levels observed in untreated controls; (A) Biokinetics after administering a single dose to male rats; (B) Biokinetics after administering a single dose to female rats; (C) Biokinetics after 14-days of daily oral treatment in male rats; (D) Biokinetics after 14-days of daily oral treatment in female rats; (E) Tissue distribution in male rats; (F) Tissue distribution in female rats. (a) and (b) is used when there is no significant difference between two groups (control and N-Cal/N-Cal and B-Cal), but significant difference is found when three groups (control, N-Cal, B-Cal) are compared.Abbreviations: B-Cal, bulk calcium carbonates; N-Cal, nano calcium carbonates.](/cms/asset/1f2aa36c-b3d4-4280-ba04-66064b764d7c/dijn_a_79403_f0003_b.jpg)
Figure S1 XRD patterns (A) and FT-IR spectra (B) of (a) B-Cal and (b) N-Cal.
Abbreviations: arb, arbitrary; B-Cal, bulk calcium carbonates; FT-IR, Fourier transform infrared; N-Cal, nano calcium carbonates; XRD, X-ray diffraction.
![Figure S1 XRD patterns (A) and FT-IR spectra (B) of (a) B-Cal and (b) N-Cal.Abbreviations: arb, arbitrary; B-Cal, bulk calcium carbonates; FT-IR, Fourier transform infrared; N-Cal, nano calcium carbonates; XRD, X-ray diffraction.](/cms/asset/e06d20d7-cb2a-453c-9687-5073d53d0476/dijn_a_79403_sf0001_b.jpg)
Figure S2 SEM images for the produced B-Cal (A, C) and N-Cal (B, D).
Abbreviations: B-Cal, bulk calcium carbonates; N-Cal, nano calcium carbonates; SEM, scanning electron microscopy.
![Figure S2 SEM images for the produced B-Cal (A, C) and N-Cal (B, D).Abbreviations: B-Cal, bulk calcium carbonates; N-Cal, nano calcium carbonates; SEM, scanning electron microscopy.](/cms/asset/27d35f44-cf9b-4a85-99b9-a6ecb3ba3df4/dijn_a_79403_sf0002_b.jpg)
Figure S3 Hydrodynamic size distributions of B-Cal (A), and N-Cal (B) in aqueous suspension as determined by dynamic light scattering.
Abbreviations: B-Cal, bulk calcium carbonates; N-Cal, nano calcium carbonates; PDI, polydispersity index
![Figure S3 Hydrodynamic size distributions of B-Cal (A), and N-Cal (B) in aqueous suspension as determined by dynamic light scattering.Abbreviations: B-Cal, bulk calcium carbonates; N-Cal, nano calcium carbonates; PDI, polydispersity index](/cms/asset/ce2998ab-6f82-40b0-b731-3a86658c9c84/dijn_a_79403_sf0003_b.jpg)
Figure S4 Zeta potential distributions of B-Cal (A), and N-Cal (B).
Abbreviations: arb, arbitrary; B-Cal, bulk calcium carbonates; N-Cal, nano calcium carbonates.
![Figure S4 Zeta potential distributions of B-Cal (A), and N-Cal (B).Abbreviations: arb, arbitrary; B-Cal, bulk calcium carbonates; N-Cal, nano calcium carbonates.](/cms/asset/9acd2f34-ca8c-4edd-97cd-bf2a0ccd3f35/dijn_a_79403_sf0004_b.jpg)
Figure S5 XRD patterns of B-Cal (a), and N-Cal (b), incubated in simulated body fluids: gastric fluid (A), intestinal fluid (B), and plasma (C) after 24 or 48 hours.
Note: Asterisks indicate the evolution of dicalcium phosphate dihydrate (JCPDS No 72-0714).
Abbreviations: arb, arbitrary; B-Cal, bulk calcium carbonates; JCPDS, Joint Committee on Powder Diffraction Standards; N-Cal, nano calcium carbonates; XRD, X-ray diffraction; h, hours.
![Figure S5 XRD patterns of B-Cal (a), and N-Cal (b), incubated in simulated body fluids: gastric fluid (A), intestinal fluid (B), and plasma (C) after 24 or 48 hours.Note: Asterisks indicate the evolution of dicalcium phosphate dihydrate (JCPDS No 72-0714).Abbreviations: arb, arbitrary; B-Cal, bulk calcium carbonates; JCPDS, Joint Committee on Powder Diffraction Standards; N-Cal, nano calcium carbonates; XRD, X-ray diffraction; h, hours.](/cms/asset/26cf2da0-c919-482e-9bf3-adf518d2f530/dijn_a_79403_sf0005_b.jpg)
Figure S6 XANES at calcium K-edge for B-Cal (A), and N-Cal (B), treated with simulated body fluid, untreated (a) and gastric fluid (b), intestinal fluid (c) and plasma (d) for 48 hours.
Note: The inset table shows the edge position obtained by secondary differentiation of peak.
Abbreviations: arb, arbitrary; B-Cal, bulk calcium carbonates; N-Cal, nano calcium carbonates; XANES, X-ray absorption near edge structure.
![Figure S6 XANES at calcium K-edge for B-Cal (A), and N-Cal (B), treated with simulated body fluid, untreated (a) and gastric fluid (b), intestinal fluid (c) and plasma (d) for 48 hours.Note: The inset table shows the edge position obtained by secondary differentiation of peak.Abbreviations: arb, arbitrary; B-Cal, bulk calcium carbonates; N-Cal, nano calcium carbonates; XANES, X-ray absorption near edge structure.](/cms/asset/867d150b-6838-4865-a70b-f56f05d10f29/dijn_a_79403_sf0006_b.jpg)
Figure S7 XRD patterns of B-Cal (a) and N-Cal (b) incubated ex vivo in fluids extracted from tissues: gastric fluid (A), intestinal fluid (B), and plasma (C).
Note: Asterisks indicate the evolution of dicalcium phosphate dihydrate (JCPDS No 72-0714).
Abbreviations: arb, arbitrary; B-Cal, bulk calcium carbonates; JCPDS, Joint Committee on Powder Diffraction Standards; N-Cal, nano calcium carbonates; XRD, X-ray diffraction; h, hours.
![Figure S7 XRD patterns of B-Cal (a) and N-Cal (b) incubated ex vivo in fluids extracted from tissues: gastric fluid (A), intestinal fluid (B), and plasma (C).Note: Asterisks indicate the evolution of dicalcium phosphate dihydrate (JCPDS No 72-0714).Abbreviations: arb, arbitrary; B-Cal, bulk calcium carbonates; JCPDS, Joint Committee on Powder Diffraction Standards; N-Cal, nano calcium carbonates; XRD, X-ray diffraction; h, hours.](/cms/asset/0b926fdd-27ea-42c6-849e-a860bf0e7d49/dijn_a_79403_sf0007_b.jpg)
![Figure S7 XRD patterns of B-Cal (a) and N-Cal (b) incubated ex vivo in fluids extracted from tissues: gastric fluid (A), intestinal fluid (B), and plasma (C).Note: Asterisks indicate the evolution of dicalcium phosphate dihydrate (JCPDS No 72-0714).Abbreviations: arb, arbitrary; B-Cal, bulk calcium carbonates; JCPDS, Joint Committee on Powder Diffraction Standards; N-Cal, nano calcium carbonates; XRD, X-ray diffraction; h, hours.](/cms/asset/749a7aa4-1cfe-4828-ab2f-8ffed0293bc9/dijn_a_79403_sf0007a_b.jpg)
Figure S8 SEM images of B-Cal (a) and N-Cal (b) after incubation ex vivo in tissue extracted fluids: gastric fluid for 1 hour (A), intestinal fluid for 4 hours (B), and plasma for 4 hours (C).
Abbreviations: B-Cal, bulk calcium carbonates; N-Cal, nano calcium carbonates; SEM, scanning electron microscopy.
![Figure S8 SEM images of B-Cal (a) and N-Cal (b) after incubation ex vivo in tissue extracted fluids: gastric fluid for 1 hour (A), intestinal fluid for 4 hours (B), and plasma for 4 hours (C).Abbreviations: B-Cal, bulk calcium carbonates; N-Cal, nano calcium carbonates; SEM, scanning electron microscopy.](/cms/asset/06d70616-efa8-43c0-9b0f-a6137f24f3b2/dijn_a_79403_sf0008_b.jpg)
![Figure S8 SEM images of B-Cal (a) and N-Cal (b) after incubation ex vivo in tissue extracted fluids: gastric fluid for 1 hour (A), intestinal fluid for 4 hours (B), and plasma for 4 hours (C).Abbreviations: B-Cal, bulk calcium carbonates; N-Cal, nano calcium carbonates; SEM, scanning electron microscopy.](/cms/asset/c6377dca-e256-4125-9dc6-4a4603972d4a/dijn_a_79403_sf0008a_b.jpg)
Figure S9 XRD patterns of B-Cal (a), and N-Cal (b), obtained from stomach (A), and intestine (B) following oral administration to rats.
Note: Asterisks and triangles indicate dicalcium phosphate dihydrate (JCPDS No 72-0714) and calcite, respectively.
Abbreviations: arb, arbitrary; B-Cal, bulk calcium carbonates; JCPDS, Joint Committee on Powder Diffraction Standards; N-Cal, nano calcium carbonates; XRD, X-ray diffraction; h, hours.
![Figure S9 XRD patterns of B-Cal (a), and N-Cal (b), obtained from stomach (A), and intestine (B) following oral administration to rats.Note: Asterisks and triangles indicate dicalcium phosphate dihydrate (JCPDS No 72-0714) and calcite, respectively.Abbreviations: arb, arbitrary; B-Cal, bulk calcium carbonates; JCPDS, Joint Committee on Powder Diffraction Standards; N-Cal, nano calcium carbonates; XRD, X-ray diffraction; h, hours.](/cms/asset/d20def11-ef4b-4e4e-afc3-29e937f4a9c2/dijn_a_79403_sf0009_b.jpg)