Abstract：
BACKGROUND: Current clinical approaches to regenerate temporomandibular joint (TMJ) articulating cartilage defects only treat the symptoms (i.e. pain and dysfunction) and do not seek to restore joint integrity for long-term relief. Therefore, we investigated a novel self-assembling tissue-engineered cartilage to overcome this significant clinical issue for TMJ regenerative purposes.
OBJECTIVE: Examine the maturation of dynamic self-regenerating cartilage (dSRC) using auricular chondrocytes and evaluate a novel combinatorial approach with fractional laser treatment and dSRC implantation for TMJ cartilage repair.
METHODS: A suspension of 107 freshly harvested rabbit ear chondrocytes was cultured under a continuous reciprocating motion to form the dSRC. After 2, 4 and 8 weeks of culture, dSRC samples were stained with H&E, Safranin-O and Toluidine Blue. Immunohistochemistry (IHC) was performed for collagens type I and II. Channels (300-500 μm diameter and 1.2-1.5 mm depth) were created in six freshly harvested condyles using a fractional Erbium laser. Two groups were tested: dSRC in a laser-ablated lesion (experimental) and an empty laser-ablated channel (control). TMJ condyles were cultured for up to 8 weeks and analysed as described above.
RESULTS: H&E staining showed a high cell density in dSRC compared to native cartilage. All dSRC groups demonstrated intense Safranin-O staining, indicating high glycosaminoglycan (GAG) production and intense Toluidine Blue staining showed high proteoglycan content. IHC confirmed that dSRC consisted predominantly of collagen type II. The experimental group showed improved cartilage repair at both time points compared to the empty channels.
CONCLUSIONS: dSRC viability and successful matrix formation were demonstrated in vitro. The combination of fractional laser ablation and dSRC implantation enhanced cartilage repair.
OBJECTIVE: Examine the maturation of dynamic self-regenerating cartilage (dSRC) using auricular chondrocytes and evaluate a novel combinatorial approach with fractional laser treatment and dSRC implantation for TMJ cartilage repair.
METHODS: A suspension of 107 freshly harvested rabbit ear chondrocytes was cultured under a continuous reciprocating motion to form the dSRC. After 2, 4 and 8 weeks of culture, dSRC samples were stained with H&E, Safranin-O and Toluidine Blue. Immunohistochemistry (IHC) was performed for collagens type I and II. Channels (300-500 μm diameter and 1.2-1.5 mm depth) were created in six freshly harvested condyles using a fractional Erbium laser. Two groups were tested: dSRC in a laser-ablated lesion (experimental) and an empty laser-ablated channel (control). TMJ condyles were cultured for up to 8 weeks and analysed as described above.
RESULTS: H&E staining showed a high cell density in dSRC compared to native cartilage. All dSRC groups demonstrated intense Safranin-O staining, indicating high glycosaminoglycan (GAG) production and intense Toluidine Blue staining showed high proteoglycan content. IHC confirmed that dSRC consisted predominantly of collagen type II. The experimental group showed improved cartilage repair at both time points compared to the empty channels.
CONCLUSIONS: dSRC viability and successful matrix formation were demonstrated in vitro. The combination of fractional laser ablation and dSRC implantation enhanced cartilage repair.
摘要：
背景:目前再生颞下颌关节(TMJ)关节软骨缺损的临床方法仅治疗症状(即疼痛和功能障碍),而不寻求恢复关节完整性以长期缓解。因此,我们研究了一种新型的自组装组织工程软骨，以克服TMJ再生的重要临床问题。
目的：使用耳廓软骨细胞检查动态自我再生软骨（dSRC）的成熟，并评估一种新型的组合方法，包括点阵激光治疗和dSRC植入用于TMJ软骨修复。
方法：在连续往复运动下培养107个新鲜收获的兔耳软骨细胞的悬浮液，以形成dSRC。培养2、4和8周后，dSRC样品用H&E染色，Safranin-O和甲苯胺蓝。对I型和II型胶原进行免疫组织化学(IHC)。使用分数铒激光在六个新收获的髁中产生通道(300-500μm直径和1.2-1.5mm深度)。测试了两组：激光消融病变中的dSRC（实验）和空的激光消融通道（对照）。将TMJ髁培养长达8周,并如上所述进行分析。
结果:H&E染色显示与天然软骨相比dSRC中的高细胞密度。所有dSRC组表现出强烈的Safranin-O染色，表明高糖胺聚糖（GAG）产量和强烈的甲苯胺蓝染色显示高蛋白聚糖含量。IHC证实dSRC主要由II型胶原组成。与空通道相比，实验组在两个时间点显示出改善的软骨修复。
结论:体外证明了dSRC活力和成功的基质形成。点阵激光消融和dSRC植入的结合增强了软骨修复。
目的：使用耳廓软骨细胞检查动态自我再生软骨（dSRC）的成熟，并评估一种新型的组合方法，包括点阵激光治疗和dSRC植入用于TMJ软骨修复。
方法：在连续往复运动下培养107个新鲜收获的兔耳软骨细胞的悬浮液，以形成dSRC。培养2、4和8周后，dSRC样品用H&E染色，Safranin-O和甲苯胺蓝。对I型和II型胶原进行免疫组织化学(IHC)。使用分数铒激光在六个新收获的髁中产生通道(300-500μm直径和1.2-1.5mm深度)。测试了两组：激光消融病变中的dSRC（实验）和空的激光消融通道（对照）。将TMJ髁培养长达8周,并如上所述进行分析。
结果:H&E染色显示与天然软骨相比dSRC中的高细胞密度。所有dSRC组表现出强烈的Safranin-O染色，表明高糖胺聚糖（GAG）产量和强烈的甲苯胺蓝染色显示高蛋白聚糖含量。IHC证实dSRC主要由II型胶原组成。与空通道相比，实验组在两个时间点显示出改善的软骨修复。
结论:体外证明了dSRC活力和成功的基质形成。点阵激光消融和dSRC植入的结合增强了软骨修复。
