Cen Yu-Jie, You De-Bo, Wang Wei, Feng Yun
Department of Ophthalmology, Peking University Third Hospital, Beijing 100191, China.
Beijing Key Laboratory of Restoration of Damaged Ocular Nerve, Peking University Third Hospital, Beijing 100191, China.
Int J Ophthalmol. 2021 Jan 18;14(1):10-18. doi: 10.18240/ijo.2021.01.02. eCollection 2021.
To construct a competent corneal lamellar substitute in order to alleviate the shortage of human corneal donor.
Rabbit mesenchymal stem cells (MSCs) were isolated from bone marrow and identified by flow cytometric, osteogenic and adipogenic induction. Xenogenic decellularized corneal matrix (XDCM) was generated from dog corneas. MSCs were seeded and cultured on XDCM to construct the tissue-engineered cornea. Post-transplantation biocompatibility of engineered corneal graft were tested by animal experiment. Rabbits were divided into two groups then underwent lamellar keratoplasty (LK) with different corneal grafts: 1) XDCM group (=5): XDCM; 2) XDCM-MSCs groups (=4): tissue-engineered cornea made up with XDCM and MSCs. The ocular surface recovery procedure was observed while corneal transparency, neovascularization and epithelium defection were measured and compared. on focal exam was performed 3mo postoperatively.
Rabbit MSCs were isolated and identified. Flow cytometry demonstrated isolated cells were CD90 positive and CD34, CD45 negative. Osteogenic and adipogenic induction verified their multipotent abilities. MSC-XDCM grafts were constructed and observed. transplantation showed the neovascularization in XDCM-MSC group was much less than that in XDCM group postoperatively. Post-transplant 3-month confocal test showed less nerve regeneration and bigger cell-absent area in XDCM-MSC group.
This study present a novel corneal tissue-engineered graft that could reduce post-operatively neovascularization and remain transparency, meanwhile shows that co-transplantation of MSCs may help increase corneal transplantation successful rate and enlarge the source range of corneal substitute to overcome cornea donor shortage.
构建一种合格的角膜板层替代物，以缓解人类角膜供体短缺的问题。
从兔骨髓中分离间充质干细胞（MSCs），并通过流式细胞术、成骨和成脂诱导进行鉴定。从犬角膜制备异种脱细胞角膜基质（XDCM）。将MSCs接种于XDCM上进行培养以构建组织工程角膜。通过动物实验检测工程化角膜移植物移植后的生物相容性。将兔分为两组，然后用不同的角膜移植物进行板层角膜移植（LK）：1）XDCM组（n = 5）：XDCM；2）XDCM-MSCs组（n = 4）：由XDCM和MSCs组成的组织工程角膜。观察眼表恢复过程，同时测量并比较角膜透明度、新生血管形成和上皮缺损情况。术后3个月进行眼部局部检查。
分离并鉴定了兔MSCs。流式细胞术显示分离的细胞CD90阳性，CD34、CD45阴性。成骨和成脂诱导证实了它们的多能性。构建并观察了MSC-XDCM移植物。移植后显示，术后XDCM-MSC组的新生血管形成明显少于XDCM组。移植后3个月的共聚焦检查显示，XDCM-MSC组的神经再生较少，无细胞区域较大。
本研究提出了一种新型的角膜组织工程移植物，可减少术后新生血管形成并保持透明度，同时表明MSCs的共移植可能有助于提高角膜移植成功率，扩大角膜替代物的来源范围以克服角膜供体短缺问题。
