Gelatin-methacrylamide gel loaded with microspheres to deliver GDNF in bilayer collagen conduit promoting sciatic nerve growth

• Published in: International journal of nanomedicine Vol. 11; no. default; pp. 1383 - 1394
• Main Authors: Zhuang, Hai, Bu, Shoushan, Hua, Lei, Darabi, Mohammad A, Cao, Xiaojian, Xing, Malcolm
• Format: Journal Article
• Language: English
• Published: New Zealand Dove Medical Press Limited 01.04.2016; Dove Medical Press
• Subjects: Acrylamides - chemistry; Animals; biodegradable nerve conduit; Collagen; Collagen - chemistry; controlled release of GDNF; Enzyme-Linked Immunosorbent Assay; Enzymes; Gelatin - chemistry; gelatin microspheres; gelatin-methacrylamide hydrogel; Glial Cell Line-Derived Neurotrophic Factor - administration & dosage; Glial Cell Line-Derived Neurotrophic Factor - metabolism; Immunoenzyme Techniques; Male; Microspheres; Nerve Regeneration - drug effects; Original Research; Rats; Rats, Sprague-Dawley; Sciatic Nerve - drug effects; Sciatic Nerve - growth & development; sciatic nerve regeneration; biodegradable nerve conduit; controlled release of GDNF; sciatic nerve regeneration; gelatin microspheres; gelatin-methacrylamide hydrogel
• ISSN: 1178-2013; 1176-9114; 1178-2013
• DOI: 10.2147/IJN.S96324

In this study, we fabricated glial cell-line derived neurotrophic factor (GDNF)-loaded microspheres, then seeded the microspheres in gelatin-methacrylamide hydrogel, which was finally integrated with the commercial bilayer collagen membrane (Bio-Gide(®)). The novel composite of nerve conduit was employed to bridge a 10 mm long sciatic nerve defect in a rat. GDNF-loaded gelatin microspheres had a smooth surface with an average diameter of 3.9±1.8 μm. Scanning electron microscopy showed that microspheres were uniformly distributed in both the GelMA gel and the layered structure. Using enzyme-linked immunosorbent assay, in vitro release studies (pH 7.4) of GDNF from microspheres exhibited an initial burst release during the first 3 days (18.0%±1.3%), and then, a prolonged-release profile extended to 32 days. However, in an acidic condition (pH 2.5), the initial release percentage of GDNF was up to 91.2%±0.9% within 4 hours and the cumulative release percentage of GDNF was 99.2%±0.2% at 48 hours. Then the composite conduct was implanted in a 10 mm critical defect gap of sciatic nerve in a rat. We found that the nerve was regenerated in both conduit and autograft (AG) groups. A combination of electrophysiological assessment and histomorphometry analysis of regenerated nerves showed that axonal regeneration and functional recovery in collagen tube filled with GDNF-loaded microspheres (GM + CT) group were similar to AG group (P>0.05). Most myelinated nerves were matured and arranged densely with a uniform structure of myelin in a neat pattern along the long axis in the AG and GM + CT groups, however, regenerated nerve was absent in the BLANK group, left the 10 mm gap empty after resection, and the nerve fiber exhibited a disordered arrangement in the collagen tube group. These results indicated that the hybrid system of bilayer collagen conduit and GDNF-loaded gelatin microspheres combined with gelatin-methacrylamide hydrogels could serve as a new biodegradable artificial nerve guide for nerve tissue engineering.