Injectable Hyaluronan/Hydroxyapatite Composite: Preparation, Physicochemical Property and Biocompatibility

doi:10.15541/jim20210020

Abstract

Abstract:

With good cytocompatibility, bioactivity, and slow degradation rate, hydroxyapatite (HAP) may remedy drawbacks of hyaluronan (HA) as soft tissue filler. Here, we prepared HA-HAP composite hydrogels by one-step reaction method, investigated various processing parameters on their physicochemical properties, and evaluated their biological performance. As-prepared HA-HAP composite hydrogels contained 1.5%-3.0% HA and 2.38%-15.8% HAP with good sterilization stability. Swelling-ratio and mechanical performance of the composite hydrogels could be adjusted by controlling HAP content and crosslinking reagent. The equilibrium swelling ratio achieved over 90% only after 1 h swell, and the swelling properties fitted well with 1^st-order exponential decay equations. By 1500 U/mL HAase solutions, the composites degraded 95% after only 2 min and dispersed rapidly, showing good enzymatic degradability. HAP was demonstrated not only endowing the composite hydrogels with better plasticity for facial soft tissue filler and contour modification, but also providing good microenvironment for attachment and proliferation of fibroblasts. Therefore, the as-prepared HA-HAP composite could be used as a long-acting filler for remodeling extracellular matrix, and thus potentially applied in cosmetic medicine.

Key words: injectable composite, hyaluronan, hydroxyapatite, physicochemical property, cytocompatibility

CLC Number:

TQ174

ZHU Yutong, TAN Peijie, LIN Hai, ZHU Xiangdong, ZHANG Xingdong. Injectable Hyaluronan/Hydroxyapatite Composite: Preparation, Physicochemical Property and Biocompatibility[J]. Journal of Inorganic Materials, 2021, 36(9): 981-990.

Figures/Tables 13

Fig. 1 Diagram of reaction between HA and BDDE

Table 1 Amount of HA, HAP and BDDE in preparing composite hydrogels

Sample	HA/wt%	HAP/wt%	BDDE equivalent
HA-1.0	10	0	1.0
HAP15-0.5	10	15	0.5
HAP30-0.5	10	30	0.5
HAP30-1.0	10	30	1.0
HAP45-1.0	10	45	1.0

Table 2 Contents of HA and HAP in different composites before and after dialysis

Sample	Before dialysis		After dialysis
Sample	HA/%	HAP/%	HA/%	HAP/%
HA-1.0	10	0	2.01	-
HAP15-0.5	10	15	1.58	2.38
HAP30-0.5	10	30	1.54	4.63
HAP30-1.0	10	30	2.19	7.73
HAP45-1.0	10	45	3.00	15.8

Fig. 2 Appearance of composite hydrogels before and after autoclaving (A) Fully dialyzed; (B) Not dialyzed. From left to right: HA-1.0, HAP15-0.5, HAP30-0.5, HAP30-1.0, and HAP45-1.0

Fig. 3 Appearance of composite hydrogels HAP45-1.0 with remained crosslinking reagents From left to right: fully dialyzed, alkali-contained, cross linker-contained

Fig. 4 Storage modulus (a), loss modulus (b) and composite viscosity (c) of hydrogels before and after sterilization

Fig. 5 SEM morphologies of hydrogels

Fig. 6 Push-out strain-stress curves of hydrogels

Fig. 7 Swelling behavior of composite hydrogel HAP45-1.0 and stabilization in vitro (A) Mass and volume swelling rate; (B) Sizes of samples before and after swelling for 8 h; (C) Stability of the samples immersed in PBS for different periods

Table 3 The 1st order exponential decay equation fitting results of hydrogel HAP45-1.0 swelling behavior

	A	y₀	R²
Mass swelling rate	-51.64	51.64	0.9992
Volume swelling rate	-78.81	78.66	0.9984

Fig. 8 Enzymatic degradation of composite hydrogel HAP45-1.0 in vitro by HAase at 120 U/mL

Fig. 9 Fibroblast adhesion after being seeded on the surface of hydrogels observed by confocal fluorescence microscope

Fig. 10 Proliferation of fibroblasts on surface of hydrogels (*p<0.05)

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