Department of Hand-, Plastic and Reconstructive Surgery, Burn Trauma Center, BG Trauma Center Ludwigshafen; University of Heidelberg, Ludwigshafen, Germany.
Department of Radiology and Nuclear Medicine, University Medical Center Mannheim, Medical Faculty Mannheim-Heidelberg University, Mannheim, Germany.
PLoS One. 2022 Jul 6;17(7):e0270112. doi: 10.1371/journal.pone.0270112. eCollection 2022.
The collagenase of the bacterium Clostridium histolyticum (CCH) is already an established treatment for fibroproliferative diseases like M. Dupuytren and M. Peyronie Although results are comparable to surgical intervention, skin laceration is a severe and relevant side effect. Doxycycline (DOX) recently rose interest as an inhibitor of matrix-metalloproteinases alongside its capabilities of skin accumulation. It therefore might be a potential skin protective agent in the use of CCH.
For simulation of a fibroproliferative disease adjacent to the skin, we utilized a rodent model of capsular fibrosis involving silicone implants and subsequent fibrotic capsule formation. For in-vitro studies, fibrotic capsules were excised and incubated with 0.9 mg/ml CCH and four different doses of DOX. For in-vivo experiments, animals received 0.0, 0.3 or 0.9 mg/ml CCH injections into the fibrotic capsules with or without prior oral DOX administration. Outcome analysis included histology, immunohistochemistry, gene expression analysis, chemical collagen and DOX concentration measurements as well as μCT imaging.
In-vitro, DOX showed a dose-dependent inhibition of CCH activity associated with increasing capsule thickness and collagen density and content. In-vivo, oral DOX administration did neither interfere with capsule formation nor in effectiveness of CCH dissolving fibrotic capsule tissue. However, skin thickness and especially collagen density was significantly higher compared to control groups. This led to a reduced rate of clinical skin lacerations after DOX administration.
DOX inhibits CCH and accumulates in the skin. Thereby, DOX can effectively reduce skin laceration after CCH treatment.
溶组织梭菌（CCH）的胶原酶已经是纤维增生性疾病（如 M. Dupuytren 和 M. Peyronie）的既定治疗方法。尽管结果与手术干预相当，但皮肤裂伤是一种严重且相关的副作用。多西环素（DOX）最近因其抑制基质金属蛋白酶的能力以及皮肤积累的能力而引起关注。因此，它可能是 CCH 治疗中潜在的皮肤保护剂。
为了模拟紧邻皮肤的纤维增生性疾病，我们利用了一种涉及硅胶植入物和随后纤维囊形成的胶囊纤维化的啮齿动物模型。对于体外研究，切除纤维囊并与 0.9mg/ml CCH 和四种不同剂量的 DOX 孵育。对于体内实验，动物接受 0.0、0.3 或 0.9mg/ml CCH 注射到纤维囊内，同时或不事先给予口服 DOX 治疗。结果分析包括组织学、免疫组织化学、基因表达分析、化学胶原和 DOX 浓度测量以及 μCT 成像。
在体外，DOX 表现出与胶囊厚度和胶原密度和含量增加相关的 CCH 活性的剂量依赖性抑制。在体内，口服 DOX 给药既不干扰囊形成，也不影响 CCH 溶解纤维囊组织的效果。然而，与对照组相比，皮肤厚度，尤其是胶原密度明显更高。这导致 DOX 给药后临床皮肤裂伤的发生率降低。
DOX 抑制 CCH 并在皮肤中积累。因此，DOX 可以有效减少 CCH 治疗后的皮肤裂伤。
