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Research Papers: Engineering Cell Microenvironment Using Novel Hydrogels

Ultraviolet-Crosslinkable and Injectable Chitosan/Hydroxyapatite Hybrid Hydrogel for Critical Size Calvarial Defect Repair In Vivo

[+] Author and Article Information
Baoqiang Li

Institute for Advanced Ceramics,
State Key Laboratory of Urban Water
Resource and Environment,
Harbin Institute of Technology,
Harbin 150001, China
e-mail: libq@hit.edu.cn

Lei Wang

Institute for Advanced Ceramics,
State Key Laboratory of Urban Water
Resource and Environment,
Harbin Institute of Technology,
Harbin 150001, China
e-mail: violetgo@126.com

Yu Hao

Institute for Advanced Ceramics,
State Key Laboratory of Urban Water
Resource and Environment,
Harbin Institute of Technology,
Harbin 150001, China
e-mail: 595750041@qq.com

Daqing Wei

Institute for Advanced Ceramics,
Harbin Institute of Technology,
Harbin 150001, China
e-mail: daqingwei@hit.edu.cn

Ying Li

Sino-Russian Institute of Hard Tissue
Development and Regeneration,
The Second Affiliated Hospital of
Harbin Medical University,
Harbin 150001, China
e-mail: Bonnieli1122@gmail.com

Yujie Feng

State Key Laboratory of Urban Water
Resource and Environment,
Harbin Institute of Technology,
Harbin 150001, China
e-mail: yujief@hit.edu.cn

Dechang Jia

Institute for Advanced Ceramics,
Harbin Institute of Technology,
Harbin 150001, China
e-mail: dcjia@hit.edu.cn

Yu Zhou

Institute for Advanced Ceramics,
Harbin Institute of Technology,
Harbin 150001, China
e-mail: zhouyu@hit.edu.cn

Manuscript received September 15, 2015; final manuscript received February 3, 2016; published online April 8, 2016. Assoc. Editor: Feng Xu.

J. Nanotechnol. Eng. Med 6(4), 041001 (Apr 08, 2016) (6 pages) Paper No: NANO-15-1080; doi: 10.1115/1.4032902 History: Received September 15, 2015; Revised February 03, 2016

To promote bone regeneration in vivo using critical-size calvarial defect model, hybrid hydrogel was prepared by mixing chitosan with hydroxyapatite (HA) and ultraviolet (UV) irradiation in situ. The hydrosoluble, UV-crosslinkable and injectable N-methacryloyl chitosan (N-MAC) was synthesized via single-step N-acylation reaction. The chemical structure was confirmed by 1H-NMR and FTIR spectroscopy. N-MAC hydrogel presented a microporous structure with pore sizes ranging from 10 to 60 μm. Approximately 80% cell viability of N-MAC hydrogel against encapsulated 3T3 cell indicated that N-MAC is an emerging candidate for mimicking native extracellular matrix (ECM). N-MAC hydrogel hybridized with HA was used to accelerate regeneration of calvarial bone using rabbit model. The effects of hybrid hydrogels to promote bone regeneration were evaluated using critical size calvarial bone defect model. The healing effects of injectable hydrogels with/without HA for bone regeneration were investigated by analyzing X-ray image after 4 or 6 weeks. The results showed that the regenerated new bone for N-MAC 100 was significantly greater than N-MAC without HA and untreated controls. The higher HA content in N-MAC/HA hybrid hydrogel benefited the acceleration of bone regeneration. About 50% closure of defect site after 6 weeks postimplantation demonstrated potent osteoinductivity of N-MAC 100 UV-crosslinkable and injectable N-MAC/HA hybrid hydrogel would allow serving as a promising biomaterial for bone regeneration using the critical-size calvarial defect.

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Figures

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Fig. 1

Synthesis, characterization, and property of UV-crosslinkable and injectable N-MAC. (a) The N-acylation scheme between chitosan and MA; (b) 1H NMR spectra (400 MHz, D2O) of N-MAC; (c) FTIR spectra of chitosan, N-MAC, and N-MAC/HA; (d) photos of N-MAC solution in inclined vial (up); N-MAC solution could form hydrogel that exhibited fixed shape, respectively, in inverted vial (down) after UV irradiation; (e) SRs of N-MAC hydrogel with DS of 28.4%; ESEM photograph of N-MAC hydrogel; (f) the XRD patterns of chitosan, N-MAC and N-MAC/HA. (g) Degradation of N-MAC and N-MAC/HA hydrogels in PBS containing 1 mg/mL papain at 37 °C.

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Fig. 2

The biocompatibility of N-MAC solution and N-MAC hydrogel. (a) NIH/3T3 cells viability of N-MAC solution with different concentrations for culture 12 hrs and culture medium was used as control group. Each point is presented as mean±standard deviation (SD) (n = 5, *p < 0.01); (b) proliferation of NIH/3T3 cells in the presence of N-MAC solution for culture 12, 24, and 36 hrs. Each point is presented as mean±SD (n = 5); (c) fluorescence images of 3T3 cell in N-MAC hydrogel. (d) Typical photos of N-MAC hydrogel adjacent with regional skin after 6 days. H&E stained histological images of subcutaneous tissue at 10 days.

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Fig. 3

Critical size calvarial defect repair using UV-crosslinkable and injectable hybrid hydrogel. (a) Representative X-ray images of isolated calvaria of only N-MAC hydrogel, N-MAC 20, N-MAC 40, and N-MAC 100 4 weeks (a) and 6 weeks (b); the statistical evaluations of apparent repair area of only N-MAC hydrogel, N-MAC 20, N-MAC 40 and N-MAC 100 after 4 weeks (c) and 6 weeks (d).

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