CAJ | 학술논문

目的设计并构建一种含有编织网的连续性真皮再生模板,并初步评价其生物学性能.方法采用冷冻-冻干法将聚乳酸-羟基乙酸(PLGA)编织网整合到胶原-壳聚糖支架(CCS)中,形成PLGA编织网/胶原-壳聚糖复合支架(PCCS),对比PLGA编织网、PCCS、CCS微观形态及机械强度.将PCCS及CCS分别埋植于18只SD大鼠皮下(PCCS组、CCS组,每组9只大鼠).术后1、2、4周取埋植部位皮肤标本,行组织形态学观察及免疫组织化学观察(用CD68抗体对巨噬细胞染色),蛋白质印迹法检测CD68、髓过氧化物酶(MPO)、IL-1β、IL-10的蛋白含量.对数据行单因素方差分析和t检验.结果含有编织网的PCCS具有类似CCS的三维多孔结构;PLGA编织网与PCCS的机械强度分别为(3.07±0.10)、(3.26±0.15)MPa,明显优于CCS[(0.42±0.21)MPa,F=592.3,P＜0.0001].PCCS组约在术后2周完成全层组织长入,CCS组于术后4周才实现这一过程.PCCS组、CCS组均出现巨噬细胞聚集的现象,在术后2周达到高峰,且CCS组可见更多数量的巨噬细胞浸润.术后2周,PCCS组IL-10蛋白含量高于CCS组,而CD68、MPO、IL-1β蛋白含量则低于CCS组(t=-4.06～2.89,P＜0.05或P＜0.01).结论支架的三维结构与机械强度之间的相互作用,在诱导组织再生中非常重要.PCCS连续性真皮再生模板具有优良的机械性能,合适的三维多孔结构和快速诱导组织、细胞及血管长人的潜能,作为真皮替代物具有一定应用前景.
목적 설계병구건일충함유편직망적련속성진피재생모판,병초보평개기생물학성능.방법 채용냉동-동간법장취유산-간기을산(PLGA)편직망정합도효원-각취당지가(CCS)중,형성PLGA편직망/효원-각취당복합지가(PCCS),대비PLGA편직망、PCCS、CCS미관형태급궤계강도.장PCCS급CCS분별매식우18지SD대서피하(PCCS조、CCS조,매조9지대서).술후1、2、4주취매식부위피부표본,행조직형태학관찰급면역조직화학관찰(용CD68항체대거서세포염색),단백질인적법검측CD68、수과양화물매(MPO)、IL-1β、IL-10적단백함량.대수거행단인소방차분석화t검험.결과 함유편직망적PCCS구유유사CCS적삼유다공결구;PLGA편직망여PCCS적궤계강도분별위(3.07±0.10)、(3.26±0.15)MPa,명현우우CCS[(0.42±0.21)MPa,F=592.3,P＜0.0001].PCCS조약재술후2주완성전층조직장입,CCS조우술후4주재실현저일과정.PCCS조、CCS조균출현거서세포취집적현상,재술후2주체도고봉,차CCS조가견경다수량적거서세포침윤.술후2주,PCCS조IL-10단백함량고우CCS조,이CD68、MPO、IL-1β단백함량칙저우CCS조(t=-4.06～2.89,P＜0.05혹P＜0.01).결론 지가적삼유결구여궤계강도지간적상호작용,재유도조직재생중비상중요.PCCS련속성진피재생모판구유우량적궤계성능,합괄적삼유다공결구화쾌속유도조직、세포급혈관장인적잠능,작위진피체대물구유일정응용전경.
Objective To design and construct a kind of dermal regeneration template with mesh,and to preliminarily evaluate its biological characteristics. Methods PLGA mesh was integrated into CCS with freeze-drying method for constructing PLGA mesh/CCS composite (PCCS). The micromorphologies and mechanical properties among PLGA mesh, CCS, and PCCS were compared. PCCS and CCS was respectively implanted into subcutaneous tissue of SD rats(PCCS and CCS groups, 9 rats in each group). The tissue samples were collected at post operation week (POW) 1, 2, and 4 for histopathological and immunohistochemical observation. Protein levels of CD68, MPO, IL-1 β, IL-10 were examined by Western blot, with expression of gray value. Data were processed with one-way analysis of variance and t test. Results Threedimensional porous structure of PCCS was similar to that of CCS. Mechanical property of PLGA mesh and PCCS was respectively (3.07 ±0. 10), (3.26 ±0. 15) MPa, and they were higher than that of CCS [(0.42 ±0.21) MPa, F = 592.3, P ＜ 0. 0001)]. The scaffolds were filled with newly formed tissue in PCCS group at POW 2, while those in CCS group were observed at POW 4. A large accumulation of macrophages was observed in both groups, especially at POW 2, and more macrophage infiltration was observed in CCS group. The protein level of IL-10 in PCCS group at POW 2 was obviously higher than that in CCS group, while the protein levels of CD68, MPO, IL-1 β were significantly decreased as compared with those in CCSgroup (with t value from -4.06to2.89,P ＜0.05orP ＜0.01). Conclusions PCCS has excellent mechanical property with appropriate three-dimensional porous structure. Meanwhile, it can rapidly induce formation of new tissue and vascularization, and it has a prospect of serving as a dermal substitute.