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Experimental study on allografts of amniotic epithelial cells in calcaneal tendon lesions of sheep

机译:羊跟骨肌腱损伤同种羊膜上皮细胞移植的实验研究

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摘要

An experimental protocol was designed to study the survival and behaviour of an allograft of amniotic epithelial cells (AECs) in an ovine model. The study was conducted on three healthy adult sheep. A core lesion was created in both calcaneal tendons under ultrasound (US) guidance by injecting 400 UI of Type 1A collagenase diluted in 0.6 ml saline. The AECs were obtained from a 60–80-day-old fetus and cultured under standard conditions. After 15 days of collagenase treatment, 2 × 106 AECs stained with a vital membrane fluorescent probe (PHK26) were injected under US guidance in 500 μl saline solution into the lesion of one limb. The contralateral untreated limb was used as a control. Animals were euthanatized 7 (1) and 30 (2) days later. Histological analyses performed on explanted tendons clearly demonstrate that AECs survived for at least 1 month inside the lesion without any adverse reactions. The damaged tissue of the treated tendons showed a high number of reparative cells in active proliferation that were accumulating collagen within the extracellular matrix. In addition, after 1 month, the neo-collagen began to be organized into parallel arrays of fibers oriented along the longitudinal axis of the tendon.
机译:设计了一个实验方案来研究羊模型中同种异体羊膜上皮细胞(AEC)的存活和行为。该研究是对三只健康的成年绵羊进行的。在超声(US)的指导下,通过注射在600 ml生理盐水中稀释的400 UI的1A型胶原酶,在两个跟骨肌腱中产生核心病变。 AECs来自60-80日龄的胎儿,并在标准条件下培养。胶原酶处理15天后,在US指导下,将500×l生理盐水中的2×10 6 AEC染色,并用活膜荧光探针(PHK26)注入一只患肢。对侧未治疗的肢体用作对照。 7(1)天和30(2)天后对动物实施安乐死。对植入的肌腱进行的组织学分析清楚地表明,AEC在病变内存活了至少1个月,没有任何不良反应。处理过的肌腱的受损组织显示出大量活跃繁殖的修复细胞,这些细胞在细胞外基质中积累胶原蛋白。此外,在1个月后,新胶原开始组织成平行的纤维阵列,沿着肌腱的纵轴排列。

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