Biomimetic Mineralization of Hydroxyapatite Mediated by Poly(L-glutamic acid) Hydrogels in Simulated Body Fluid†

doi:10.7503/cjcu20180718

Abstract

Abstract:

Bioactive poly(L-glutamic acid)(PLGA) hydrogels were prepared by using adipic acid dihydrazide(ADH) as a crosslinking agent. The formation and growth of hydroxyapatite(HA) in the simulated body fluid(SBF) of different concentrations were characterized by various techniques. PLGA is a kind of anionic poly(amino acid) with abundant carboxylic groups, which favors the enrichment of calcium ions and formation of HA. The growth of HA with soaking time was detected at the surface and interior of the PLGA hydrogels. The mechanical properties of the PLGA hydrogels before and after mineralization were studied in dynamic rheology experiments. Being cytocompatible, the mineralized PLGA hydrogels were successfully applied for cultivation of adipose-derived stem cells(ASCs). These results presented a method of preparing HA/PLGA composite hydrogels, which might be used as a promising biomaterial for bone tissue engineering.

Key words: Poly(L-glutamic acid) hydrogel, Biomimetic mineralization, Hydroxyapatite, Bone repair

CLC Number:

O636

TrendMD:

YAN Shifeng,WANG Weidong,REN Jie,TENG Changchang,YIN Jingbo. Biomimetic Mineralization of Hydroxyapatite Mediated by Poly(L-glutamic acid) Hydrogels in Simulated Body Fluid^†[J]. Chem. J. Chinese Universities, 2019, 40(4): 815.

Figures/Tables 13

Table 1 Comparation of the ion concentration(mmol/L) in human plasma, 1.0SBF and 1.5SBF

Sample	Ion concentration/(mmol·L^-1)
Sample	Na⁺	K⁺	Ca²⁺	Mg²⁺	HC $O 3 -$	Cl^-	HP $O 4 2 -$	S $O 4 2 -$
Plasma	142.0	5.0	2.5	1.5	27.0	103.0	1.0	0.5
1.0SBF	142.0	5.0	2.5	1.5	4.2	148.0	1.0	0.5
1.5SBF	213.1	7.5	3.8	2.3	6.3	221.9	1.5	0.8

Table 1 Comparation of the ion concentration(mmol/L) in human plasma, 1.0SBF and 1.5SBF

Sample	Ion concentration/(mmol·L^-1)
Sample	Na⁺	K⁺	Ca²⁺	Mg²⁺	HC $O 3 -$	Cl^-	HP $O 4 2 -$	S $O 4 2 -$
Plasma	142.0	5.0	2.5	1.5	27.0	103.0	1.0	0.5
1.0SBF	142.0	5.0	2.5	1.5	4.2	148.0	1.0	0.5
1.5SBF	213.1	7.5	3.8	2.3	6.3	221.9	1.5	0.8

Scheme 1 Schematic of PLGA crosslinking

Fig.1 Photographs of PLGA/ADH solution before(A) and after addition of EDC(B—D)(B) Investion; (C) lifting up; (D) picking up.

Fig.2 XRD patterns of PLGA hydrogels after soaking in 1.5SBF for different time(A) and comparison of XRD patterns of PLGA hydrogels after soaking in 1.0SBF and 1.5SBF(B)

Fig.3 FTIR spectra of PLGA(a), PLGA hydrogel before(b)and after mineralization(c)

Fig.4 Photographs of PLGA hydrogels before(A) and after soaking in 1.0SBF(B, B')/1.5SBF(C, C') for 14 d(B, C) and 28 d(B', C')

Fig.5 SEM images of the mineralized surface of PLGA hydrogels after soaking in 1.0SBF and 1.5SBF for different time (A—F) 2, 6, 10, 14, 21 and 28 d in 1.0SBF; (A'—F') 2, 6, 10, 14, 21 and 28 d in 1.5SBF; (G, H) different magnification in 1.5SBF for 14 d.

Fig.6 EDS spectrum of the cross-section of the hydrogel after soaking in 1.5SBF for 14 d

Fig.7 TGA curves of PLGA hydrogels after mineralization (A) PLGA hydrogels after soaking in 1.5SBF for different time; (B) comparison of TGA curves for PLGA hydrogels after soaking in 1.0SBF and 1.5SBF; (C) percentage of mass increase with immersion time for PLGA hydrogels after soaking in 1.0SBF and 1.5SBF.

Scheme 2 Schematic illustration of the HA mineralization induced by PLGA hydrogels Naka K(ed)

Fig.8 Rheological properties of PLGA hydrogels after mineralization(A) Storage modulus(G') or loss modulus(G″); (B) complex viscosity |η*| of PLGA hydrogels after soaking in 1.5SBF for different time.

Fig.9 SEM images of PLGA hydrogels before(A) and after soaking in 1.5SBF for 14 d(B) and the ASCs proliferated on PLGA hydrogel(C) and mineralized hydrogel for 3 d(D)

Fig.10 CLSM images of the ASCs-seeded PLGA hydrogels(A—C) and the ASCs-seeded mineralized hydrogel(D—F) at 1 d(A, D), 7 d(B, C) and 14 d(C, F) The living cells were stained with FDA(green) and the dead cell nuclei were stained with PI(red).

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