CAJ | 학술논문

目的：探讨正常和糖尿病来源间充质干细胞(MSCs)的生物学特性及对糖尿病慢性创面修复效果的差异。方法：采用高脂(高糖)饲料饲养1个月、腹腔注射40 mg/kg 链脲佐菌素连续3 d的方法制备糖尿病小鼠模型。取正常及糖尿病模型小鼠股骨和胫骨的骨髓,采用Percoll梯度离心法分离 MSCs,体外扩增,倒置相差显微镜下观察细胞形态学,油红“O”和茜素红染色观察其成脂、成骨细胞分化能力,流式细胞仪检测表面抗原表达,酶联免疫检测细胞增殖能力,凋亡试剂盒检测细胞的抗凋亡能力。16只糖尿病模型小鼠随机分为正常 MSCs治疗组和糖尿病MSCs治疗组(n=8),背部制作直径8 mm圆形全层皮肤缺损创面,分别创周皮下注射2×106个正常和糖尿病小鼠来源MSCs,观察创面愈合情况。结果：相同体外培养条件下,糖尿病小鼠来源的MSCs较正常小鼠来源的 MSCs形成细胞集落的能力差且出现生长抑制的时间早,细胞增殖分化能力弱,抗凋亡能力下降,流式细胞术结果显示CD29和CD105阳性表达下降。正常MSCs显示出更好地促进糖尿病慢性创面愈合的效应,尤其是在伤后7~14 d。结论：糖尿病自体的MSCs治疗能力下降,不是治疗糖尿病创面的最佳种子细胞。
목적：탐토정상화당뇨병래원간충질간세포(MSCs)적생물학특성급대당뇨병만성창면수복효과적차이。방법：채용고지(고당)사료사양1개월、복강주사40 mg/kg 련뇨좌균소련속3 d적방법제비당뇨병소서모형。취정상급당뇨병모형소서고골화경골적골수,채용Percoll제도리심법분리 MSCs,체외확증,도치상차현미경하관찰세포형태학,유홍“O”화천소홍염색관찰기성지、성골세포분화능력,류식세포의검측표면항원표체,매련면역검측세포증식능력,조망시제합검측세포적항조망능력。16지당뇨병모형소서수궤분위정상 MSCs치료조화당뇨병MSCs치료조(n=8),배부제작직경8 mm원형전층피부결손창면,분별창주피하주사2×106개정상화당뇨병소서래원MSCs,관찰창면유합정황。결과：상동체외배양조건하,당뇨병소서래원적MSCs교정상소서래원적 MSCs형성세포집락적능력차차출현생장억제적시간조,세포증식분화능력약,항조망능력하강,류식세포술결과현시CD29화CD105양성표체하강。정상MSCs현시출경호지촉진당뇨병만성창면유합적효응,우기시재상후7~14 d。결론：당뇨병자체적MSCs치료능력하강,불시치료당뇨병창면적최가충자세포。
Objective:To explore the biological characteristics and effects on wound healing of mesenchymal stem cells (MSCs)from normal and diabetic mouse.Methods:The diabetic model was replicated by feeding with high fat (high sugar)chow for 1 month and receiving intraperitoneal injection of 40 mg/kg streptozocin for 3 days in mice. MSCs were isolated from bone marrow of normal or diabetic mice,and they were purified by Percoll gradient cen-trifugation and cultured invitro. The growth of normal and diabetic MSCs were observed under phase contrast mi-croscope,and their potential of differentiation to adipocytes and osteocytes were observed by oil red O or alizarin red staining. The proliferation of MSCs was investigated by enzyme-linked immunoadsorbent assay. The antigens on cell surface were detected by FACS and the ability of cell anti-apoptosis was assessed by using the detection kits. Sixteen diabetic mice were randomly divided into the normal MSCs treatment group and the diabetic MSCs treatment group(n=8).Circular full-thickness wounds with the diameter of which 8 mm were created on the dor-sum of mice. The mice in the two groups received subcutaneous injection of 2×106 either normal or diabetic MSCs around the wound. The wound healing process was assessed with sequential digital photographs.Results:The MSCs from diabetic mice group showed decreased expressions of CD2 9 and CD1 0 5 by FACS,with weaker capaci-ty of forming clones,proliferation and differentiation,and with weaker growth capacity,which appeared earlier compared to those of the normal MSCs. The normal MSCs exhibited better effects in promoting process of wound healing,especially during 7-14 days after injury. Conclusions:The diabetic MSCs have poorer biological character-istics and treatment effect on wounds compared with that of the normal MSCs. The results also suggest that dia-betic MSCs are not the worthwhile seed cells in treating diabetic wounds.