Department of Ophthalmology, Guanghua Integrative Medicine Hospital, Shanghai, China.
PLoS One. 2013 Oct 1;8(10):e76103. doi: 10.1371/journal.pone.0076103. eCollection 2013.
This study was designed to explore the feasibility of using autologous rabbit adipose derived stem cells (rASCs) as seed cells and polylactic-co-glycolic acid (PLGA) as a scaffold for repairing corneal stromal defects. rASCs isolated from rabbit nape adipose tissue were expanded and seeded on a PLGA scaffold to fabricate cell-scaffold constructs. After 1 week of cultivation in vitro, the cell-scaffold complexes were transplanted into corneal stromal defects in rabbits. In vivo, the autologous rASCs-PLGA constructed corneal stroma gradually became transparent without corneal neovascularization after 12 weeks. Hematoxylin and eosin staining and transmission electron microscopy examination revealed that their histological structure and collagen fibril distribution at 24 weeks after implantation were similar to native counterparts. As to the defect treated with PLGA alone, the stromal defects remained. And scar tissue was observed in the untreated-group. The implanted autologous ASCs survived up to 24 weeks post-transplantation and differentiated into functional keratocytes, as assessed by the expression of aldehyde-3-dehydrogenase1A1 (ALDH1A1) and cornea-specific proteoglycan keratocan. Our results revealed that autologous rASCs could be one of the cell sources for corneal stromal restoration in diseased corneas or for tissue engineering of a corneal equivalent.
本研究旨在探讨以自体兔脂肪来源干细胞(rASCs)作为种子细胞和聚乳酸-羟基乙酸共聚物(PLGA)作为支架修复角膜基质缺损的可行性。从兔颈背部脂肪组织中分离出 rASCs 并扩增,然后接种到 PLGA 支架上,以构建细胞-支架复合物。体外培养 1 周后,将细胞-支架复合物移植到兔角膜基质缺损部位。在体内,经过 12 周后,自体 rASCs-PLGA 构建的角膜基质逐渐变得透明,没有角膜新生血管形成。苏木精-伊红染色和透射电镜检查显示,在植入后 24 周时,其组织学结构和胶原纤维排列与天然角膜相似。而单独使用 PLGA 治疗的基质缺损部位仍存在,未治疗组则观察到瘢痕组织。移植的自体 ASCs 在移植后 24 周内存活,并分化为功能性角膜细胞,这可以通过醛脱氢酶 1A1(ALDH1A1)和角膜特异性蛋白聚糖角膜蛋白的表达来评估。我们的结果表明,自体 rASCs 可以成为病变角膜或角膜等效物组织工程中角膜基质修复的细胞来源之一。