Microparticles for controlled growth differentiation factor 6 delivery to direct adipose stem cell-based nucleus pulposus regeneration

被引：24
作者：
Hodgkinson, Tom [1 ]
Stening, Jasmine Z. [2 ]
White, Lisa J. [2 ]
Shakesheff, Kevin M. [2 ]
Hoyland, Judith A. [1 ,3 ]
Richardson, Stephen M. [1 ]
机构：
[1] Univ Manchester, Manchester Acad Hlth Sci Ctr, Fac Biol Med & Hlth, Sch Biol Sci,Div Cell Matrix Biol & Regenerat Med, Oxford Rd, Manchester M13 9PT, Lancs, England
[2] Univ Nottingham, Fac Sci, Sch Pharm, Div Regenerat Med & Cellular Therapies, Nottingham, England
[3] NIHR Manchester Biomed Res Ctr, Manchester Acad Hlth Sci Ctr, Cent Manchester Fdn Trust, Manchester, Lancs, England
来源：
JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE | 2019年 / 13卷 / 08期
基金：
英国医学研究理事会;
关键词：
adipose stem cell; growth differentiation factor 6; intervertebral disc degeneration; microparticle; nucleus pulposus; regeneration; HUMAN INTERVERTEBRAL DISC; IN-VITRO; NEEDLE PUNCTURE; ECONOMIC BURDEN; BACK-PAIN; DEGENERATION; DEGRADATION; EXPRESSION; MANAGEMENT; MUTATIONS;
D O I：
10.1002/term.2882
中图分类号：
Q813 [细胞工程];
学科分类号：
摘要：
Currently, there is no effective long-term treatment for intervertebral disc (IVD) degeneration, making it an attractive candidate for regenerative therapies. Hydrogel delivery of adipose stem cells (ASCs) in combination with controlled release of bioactive molecules is a promising approach to halt IVD degeneration and promote regeneration. Growth differentiation factor 6 (GDF6) can induce ASC differentiation into anabolic nucleus pulposus (NP) cells and hence holds promise for IVD regeneration. Here, we optimised design of novel poly(DL-lactic acid-co-glycolic acid) (PLGA)-polyethylene glycol-PLGA microparticles to control GDF6 delivery and investigated effect of released GDF6 on human ASCs differentiation to NP cells. Recombinant human (rh)GDF6 was loaded into microparticles and total protein and rhGDF6 release assessed. The effect of microparticle loading density on distribution and gel formation was investigated through scanning electron microscopy. ASC differentiation to NP cells was examined after 14 days in hydrogel culture by quantitative polymerase chain reaction, histological, and immunohistochemical staining in normoxic and IVD-like hypoxic conditions. RhGDF6 microparticles were distributed throughout gels without disrupting gelation and controlled rhGDF6 release over 14 days. Released GDF6 significantly induced NP differentiation of ASCs, with expression comparable with or exceeding media supplemented rhGDF6. Microparticle-delivered rhGDF6 also up-regulated sulphated glycosaminoglycan and aggrecan secretion in comparison with controls. In hypoxia, microparticle-delivered rhGDF6 continued to effectively induce NP gene expression and aggrecan production. This study demonstrates the effective encapsulation and controlled delivery of rhGDF6, which maintained its activity and induced ASC differentiation to NP cells and synthesis of an NP-like matrix suggesting suitability of microparticles for controlled growth factor release in regenerative strategies for treatment of IVD degeneration.
引用
收藏
页码：1406 / 1417
页数：12
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